This document summarizes advances in seed testing technologies for major crops. It discusses the history and concepts of seed testing, including assessing genetic purity, physical purity, physiological quality, and seed health. Modern methods like molecular markers, image analysis, and spectral imaging provide non-destructive, quick, and highly accurate testing compared to traditional techniques. These advances allow for improved evaluation of seed quality attributes and performance.
Physical purity analysis tells us the proportion of pure seed component in the seed lot as well as the proportion of other crop seed, weed seed and inert matter by weight in percentage for which Seed Standards have been prescribed.
Thus, it helps in:
Improving the plant stand (by increasing the pure seed component).
Raising a pure crop (by eliminating other crop seed and weed seeds).
Raising a disease free-crop (by eliminating inert matter).
In the use of seed drill (by selecting uniform particles).
The document discusses various tests used to assess seed viability and vigor, including warm germination tests, tetrazolium tests, growth tests, and stress tests. The warm germination test is the standard test to assess viability by germinating seeds in ideal conditions. Tetrazolium tests use chemicals to determine potential germination. Growth tests measure speed and size of seedling growth. Stress tests like cold tests and accelerated aging expose seeds to stressful conditions to evaluate vigor. Proper testing helps farmers make management decisions about seeding rates and avoiding weeds.
Seed vigour is determined by the properties that allow seeds to germinate and grow in different environments. It is affected by both internal factors, like genotype and seed size, and external factors such as mechanical injury during harvesting, pre-harvesting conditions, soil temperature and moisture, tillage and fertilizer use, and moisture uptake. Smaller seeds tend to be less vigorous than medium or large seeds. Harvesting seeds before maturity and mechanical damage during harvesting can reduce seed vigour by allowing pathogens to enter seeds. High soil moisture and temperature during seed development and storage can increase respiration and pathogen growth, shortening storage life. Tillage and adequate fertilizer promote seed yield and vigour.
This document summarizes the floral biology of tomatoes. It begins with the botanical name (Solanum lycopersicum) and family (Solanaceae). It then describes the plant's roots, stem, leaves, inflorescence, flower morphology including calyx, corolla, androecium, and gynoecium. It discusses anthesis, self-pollination, cross-pollination techniques including emasculation, pollen collection, and artificial pollination. It also covers fertilization, fruit set, seed structure and composition, economic importance as a major global crop and model plant for research.
This document provides information about bitter gourd and bottle gourd. It discusses the botany, origin, domestication, breeding, and improved varieties of bitter gourd. It describes that bitter gourd is widely cultivated in many countries for its nutrient-rich immature fruits. The document also summarizes key details about the botany and origin of bottle gourd from Africa and Asia. It mentions that bottle gourd is a monoecious vine grown for its tender fruits and used to make sweets.
Cucumbers are an important crop that are a good source of potassium and various antioxidants. They prefer light, well-drained soils and temperatures between 20-28°C for optimal growth and production. Common diseases include downy mildew and powdery mildew. Management involves growing resistant varieties, maintaining good airflow, removing crop debris, and applying fungicides like mancozeb. Popular varieties in India include Pusa Sanyog, Pusa Uday, and Japanese Long Green.
Physical purity analysis tells us the proportion of pure seed component in the seed lot as well as the proportion of other crop seed, weed seed and inert matter by weight in percentage for which Seed Standards have been prescribed.
Thus, it helps in:
Improving the plant stand (by increasing the pure seed component).
Raising a pure crop (by eliminating other crop seed and weed seeds).
Raising a disease free-crop (by eliminating inert matter).
In the use of seed drill (by selecting uniform particles).
The document discusses various tests used to assess seed viability and vigor, including warm germination tests, tetrazolium tests, growth tests, and stress tests. The warm germination test is the standard test to assess viability by germinating seeds in ideal conditions. Tetrazolium tests use chemicals to determine potential germination. Growth tests measure speed and size of seedling growth. Stress tests like cold tests and accelerated aging expose seeds to stressful conditions to evaluate vigor. Proper testing helps farmers make management decisions about seeding rates and avoiding weeds.
Seed vigour is determined by the properties that allow seeds to germinate and grow in different environments. It is affected by both internal factors, like genotype and seed size, and external factors such as mechanical injury during harvesting, pre-harvesting conditions, soil temperature and moisture, tillage and fertilizer use, and moisture uptake. Smaller seeds tend to be less vigorous than medium or large seeds. Harvesting seeds before maturity and mechanical damage during harvesting can reduce seed vigour by allowing pathogens to enter seeds. High soil moisture and temperature during seed development and storage can increase respiration and pathogen growth, shortening storage life. Tillage and adequate fertilizer promote seed yield and vigour.
This document summarizes the floral biology of tomatoes. It begins with the botanical name (Solanum lycopersicum) and family (Solanaceae). It then describes the plant's roots, stem, leaves, inflorescence, flower morphology including calyx, corolla, androecium, and gynoecium. It discusses anthesis, self-pollination, cross-pollination techniques including emasculation, pollen collection, and artificial pollination. It also covers fertilization, fruit set, seed structure and composition, economic importance as a major global crop and model plant for research.
This document provides information about bitter gourd and bottle gourd. It discusses the botany, origin, domestication, breeding, and improved varieties of bitter gourd. It describes that bitter gourd is widely cultivated in many countries for its nutrient-rich immature fruits. The document also summarizes key details about the botany and origin of bottle gourd from Africa and Asia. It mentions that bottle gourd is a monoecious vine grown for its tender fruits and used to make sweets.
Cucumbers are an important crop that are a good source of potassium and various antioxidants. They prefer light, well-drained soils and temperatures between 20-28°C for optimal growth and production. Common diseases include downy mildew and powdery mildew. Management involves growing resistant varieties, maintaining good airflow, removing crop debris, and applying fungicides like mancozeb. Popular varieties in India include Pusa Sanyog, Pusa Uday, and Japanese Long Green.
This document provides information about seed germination testing methods. It defines seed germination as the budding of a seed after being planted. Seed germination testing is conducted to predict field performance, obtain planting values, and compare germination rates between seed lots. Common substrates used include paper, sand, and soil. Seeds are placed on or between the layers of these substrates in trays under controlled temperature and moisture conditions. Proper lighting, cleaning, and breaking of dormancy are also required. Germination rates are calculated based on the number of normal seedlings observed over a testing period, usually 7-14 days.
This document provides information about seed purity analysis and germination testing procedures. It discusses the steps involved in purity analysis, including removing debris, separating lightweight materials, and examining seeds for damage. It also describes how to calculate purity percentages. For germination testing, it outlines the materials needed, sample size, placement methods on filter paper or between paper towels, and incubation conditions. Evaluation involves classifying seedlings as normal or abnormal based on root and shoot development.
This document discusses two methods for producing hybrid rice: two-line and three-line breeding systems. The two-line system uses environmentally sensitive genetic male sterility (EGMS) or chemically induced male sterility (CIMS) to produce hybrid seed. EGMS lines are male sterile under certain temperature or photoperiod conditions. Two-line hybrids have advantages over three-line such as lower production costs and greater genetic diversity of parents. China has had success adopting two-line hybrid rice, which now covers over 2 million hectares. Further research aims to develop more stable EGMS lines and higher-yielding two-line hybrids with stress tolerance and quality traits.
The document discusses genetic principles of seed production and certification. It explains that varieties can deteriorate due to developmental variations, mechanical mixtures, mutations, natural crossing, minor genetic variations, diseases, and improper techniques. Seed production and certification aims to maintain genetic purity and prevent such deterioration. It involves controlling the seed source, isolation distances, rouging fields, and certification of seeds in classes from breeder to foundation to registered to certified.
Hybrid seed production is important for vegetables to harness heterosis and ensure food security. The document discusses various techniques for hybrid seed production in different vegetable crops. It describes the principles of hybrid seed production including production of inbred lines, hybridization techniques like hand emasculation and pollination, and use of male sterility. Specific techniques for tomato, eggplant, pepper, cucurbits like watermelon and pumpkin are provided detailing crop management practices and hybridization steps to produce hybrid seeds commercially.
The seed plot technique is used to produce healthy potato seed with low virus incidence. It involves selecting virus-free plants and storing their tubers separately to plant in isolated seed plots during periods of low aphid activity. This technique produces 2600 tons of breeder seed annually in India, saving $484 million by reducing seed imports. Studies show seed plot systems yield 6-15% higher than normal cultivation practices in Kenya, Uganda, and Ethiopia.
Scope Of Vegetable Seed Production Under Protected Cultivation.pptxAnusha K R
Protected cultivation provides many-fold advantages over open field seed production of vegetables. The beauty of vegetable hybrid seed production under protected conditions is that it could be implemented at a micro or macro level depending upon the need, space, and seed crop requirements. This technology is highly productive, amenable to automation, conserves water, fertilizer, and land, and provides the required environment to overcome the biotic and abiotic stress and enhance yield as well as the quality of seeds. Protected cultivation offers a very congenial environment for producing healthy, virus-free, and genetically pure hybrid seed with higher seed yield per unit area.
This document summarizes a seminar on seed quality enhancement techniques. It begins with introductions to seed quality concepts and factors that impact seed quality. It then describes various techniques used to improve seed quality, including seed hydration/priming, coating, pelleting, and encrusting. The objectives of these techniques are to reduce seeding rates, improve germination under stress, supply nutrients and protectants, and ensure uniform field establishment. The document provides details on various priming methods and materials used for coating, pelleting, and encrusting seeds.
Grafting is an ancient asexual propagation technique where the rootstock and scion from two plants are joined together to form a single plant. The production of grafted vegetable plants first began in Japan and Korea in the late 1920s with watermelons grafted onto pumpkin rootstocks. Now common in parts of Asia, Europe and the Middle East, grafting is used to improve traits like disease tolerance, abiotic stress resistance, and yield in important vegetable crops. Modern grafting methods include hole insertion grafting, tongue approach grafting and cleft grafting. Healing chambers are used to promote graft union formation and robotic grafting systems can produce hundreds of grafted plants per hour.
This document provides information on onion seed production including floral biology, methods, requirements, and standards. It discusses:
1) Onion flowers are protandrous, with pollen shed occurring before stigma receptivity. Cross-pollination by insects is needed for high seed yields.
2) The bulb-to-seed method is most common, involving selecting bulbs in one season for planting the next season for seed production. Care is taken to select true-to-type bulbs.
3) Seed production requires isolation distances between varieties, rogueing of off-types, and inspection to ensure varietal purity and high seed quality and yields.
This document discusses seed storability and viability prediction in important oilseed crops. It covers several key topics:
1) Factors that affect seed viability and storability during storage like moisture content, temperature, packaging material and storage structure. Lower moisture content and temperature helps extend seed life.
2) Methods for predicting seed viability like accelerated aging tests, mathematical models and nomographs that relate viability to moisture content and temperature over time.
3) Biochemical changes that occur during storage like lipid peroxidation and accumulation of free radicals that can damage cell membranes and organelles leading to loss of viability over time.
4) The importance of understanding these factors and developing strategies to optimize seed storage conditions and predict viability
The document discusses guidelines for releasing and notifying crop cultivars in India. It explains that releasing a cultivar makes it available for public cultivation and allows farmers to choose varieties, while notification regulates seed quality under the Seeds Act. The process involves variety evaluation through regional trials over multiple locations and years before the State and Central Variety Release Committees decide on release. Notified varieties can then be certified to ensure standard seed quality. Advantages of notification include compulsory certification for seed production and regulation of quality for seed sales. Examples of notified rice, wheat and black gram varieties in different states are also provided.
This document provides information about soybean breeding in three parts. It begins with an introduction to soybean including its scientific name, family, uses, protein and oil production. Next is a history of soybean originating in China and spreading to other countries. The document concludes with descriptions of soybean plants, their floral biology, selfing and crossing techniques, genetics of traits, and seed production and certification standards.
This document discusses sex expression in cucurbitaceous crops. It notes that cucurbits can have various sex forms ranging from hermaphroditic to monoecious to gynoecious. The major cucurbits like cucumber, bitter gourd, muskmelon, watermelon are typically monoecious. Gynoecious lines have been developed in some crops like cucumber and muskmelon through breeding. Sex determination is controlled by genes but can be modulated by environmental factors like temperature and photoperiod. Chemicals like silver nitrate and gibberellic acid can also induce staminate flowers.
This document discusses seed processing and storage. The objectives of seed processing are to improve quality by removing impurities, maintaining viability and vigor, making handling easier, and increasing value. Methods used for processing include drying, cleaning, grading, packaging, labeling, and treatment. Storage aims to preserve seeds under controlled conditions to prolong viability for long periods. Factors that affect seed longevity are seed type, quality, coat integrity, moisture content, and storage environment. Orthodox seeds can be stored long-term at low temperature and humidity while recalcitrant seeds require different storage methods.
Seed refers to a fertilized ovule containing an embryo that can develop into a new plant. Scientifically, seed is defined as a fertilized mature ovule covered by a seed coat. There are several types of propagating materials that are also considered seeds, including tubers, bulbs, rhizomes, roots, cuttings, and grafts. Seeds are classified into different categories based on their origin and intended use, including nucleus, breeder, foundation, and certified seeds, with each subsequent category representing a larger scale of multiplication while maintaining genetic and physical purity standards.
The document discusses the taxonomy, origin, evolution, and breeding of tomatoes. It notes that tomatoes originated in South America and were domesticated from wild cherry tomatoes. Important tomato breeding objectives include increasing yield, improving fruit quality and shelf life, and developing resistance to diseases and abiotic stresses. Breeding methods discussed are introduction, selection, hybridization, and biotechnology. The first genetically modified tomato, Flavr Savr, was developed to have increased shelf life but did not significantly improve firmness.
Seed processing is a vital part of ensuring high quality seed for end users. It includes cleaning, drying, treatment, packaging, and storage. The goals of seed processing are to reduce bulk, increase longevity by drying to a safe moisture level and treating with protectants, reduce variability in vigor, and improve uniformity in size and shape. The sequence of operations typically includes drying, receiving, pre-cleaning, conditioning, cleaning, separating, treating, weighing, bagging, and storage or shipping. Processing aims to separate inert materials and weed seeds from the seed lot while upgrading quality by eliminating damaged or low vigor seeds to obtain a high percentage of pure seed with maximum germination potential.
- Tomato (Lycopersicon esculentum) is the second most consumed vegetable in the world after potatoes. It is cultivated throughout temperate and tropical regions for its nutrient-rich fruit.
- Tomatoes originate from the Andean region of South America but were domesticated in Mexico. They were later spread worldwide by Europeans.
- Tomatoes have chasmogamous flowers that self-pollinate but can also cross-pollinate up to 47% depending on conditions. Manual pollination techniques include emasculation, pollen collection and application.
Traditional phenotypic methods and newer genotypic methods can both be used to identify bacteria. Phenotypic methods include gram staining, culturing, and analyzing biochemical characteristics and reactions. These methods have limitations as some bacteria cannot be cultured. Genotypic methods like MALDI-TOF, PCR, and microarrays identify bacteria based on their genetic material and can identify bacteria directly from clinical samples faster than phenotypic methods. A variety of biochemical tests are used as part of phenotypic identification to analyze carbohydrate metabolism, production of specific compounds, enzyme activity, and other characteristics.
This document discusses various methods for assessing genetic purity in plants, including morphological, chemical, and electrophoresis-based methods. Morphological methods involve examining seed or plant traits under magnification or in a grow-out test. Chemical methods analyze seed components like secondary metabolites and proteins. Electrophoresis separates proteins or DNA based on size and charge, allowing comparison of banding patterns between varieties. Together, these methods allow testing seed samples against a pure reference to validate their genetic purity or identify off-types.
This document provides information about seed germination testing methods. It defines seed germination as the budding of a seed after being planted. Seed germination testing is conducted to predict field performance, obtain planting values, and compare germination rates between seed lots. Common substrates used include paper, sand, and soil. Seeds are placed on or between the layers of these substrates in trays under controlled temperature and moisture conditions. Proper lighting, cleaning, and breaking of dormancy are also required. Germination rates are calculated based on the number of normal seedlings observed over a testing period, usually 7-14 days.
This document provides information about seed purity analysis and germination testing procedures. It discusses the steps involved in purity analysis, including removing debris, separating lightweight materials, and examining seeds for damage. It also describes how to calculate purity percentages. For germination testing, it outlines the materials needed, sample size, placement methods on filter paper or between paper towels, and incubation conditions. Evaluation involves classifying seedlings as normal or abnormal based on root and shoot development.
This document discusses two methods for producing hybrid rice: two-line and three-line breeding systems. The two-line system uses environmentally sensitive genetic male sterility (EGMS) or chemically induced male sterility (CIMS) to produce hybrid seed. EGMS lines are male sterile under certain temperature or photoperiod conditions. Two-line hybrids have advantages over three-line such as lower production costs and greater genetic diversity of parents. China has had success adopting two-line hybrid rice, which now covers over 2 million hectares. Further research aims to develop more stable EGMS lines and higher-yielding two-line hybrids with stress tolerance and quality traits.
The document discusses genetic principles of seed production and certification. It explains that varieties can deteriorate due to developmental variations, mechanical mixtures, mutations, natural crossing, minor genetic variations, diseases, and improper techniques. Seed production and certification aims to maintain genetic purity and prevent such deterioration. It involves controlling the seed source, isolation distances, rouging fields, and certification of seeds in classes from breeder to foundation to registered to certified.
Hybrid seed production is important for vegetables to harness heterosis and ensure food security. The document discusses various techniques for hybrid seed production in different vegetable crops. It describes the principles of hybrid seed production including production of inbred lines, hybridization techniques like hand emasculation and pollination, and use of male sterility. Specific techniques for tomato, eggplant, pepper, cucurbits like watermelon and pumpkin are provided detailing crop management practices and hybridization steps to produce hybrid seeds commercially.
The seed plot technique is used to produce healthy potato seed with low virus incidence. It involves selecting virus-free plants and storing their tubers separately to plant in isolated seed plots during periods of low aphid activity. This technique produces 2600 tons of breeder seed annually in India, saving $484 million by reducing seed imports. Studies show seed plot systems yield 6-15% higher than normal cultivation practices in Kenya, Uganda, and Ethiopia.
Scope Of Vegetable Seed Production Under Protected Cultivation.pptxAnusha K R
Protected cultivation provides many-fold advantages over open field seed production of vegetables. The beauty of vegetable hybrid seed production under protected conditions is that it could be implemented at a micro or macro level depending upon the need, space, and seed crop requirements. This technology is highly productive, amenable to automation, conserves water, fertilizer, and land, and provides the required environment to overcome the biotic and abiotic stress and enhance yield as well as the quality of seeds. Protected cultivation offers a very congenial environment for producing healthy, virus-free, and genetically pure hybrid seed with higher seed yield per unit area.
This document summarizes a seminar on seed quality enhancement techniques. It begins with introductions to seed quality concepts and factors that impact seed quality. It then describes various techniques used to improve seed quality, including seed hydration/priming, coating, pelleting, and encrusting. The objectives of these techniques are to reduce seeding rates, improve germination under stress, supply nutrients and protectants, and ensure uniform field establishment. The document provides details on various priming methods and materials used for coating, pelleting, and encrusting seeds.
Grafting is an ancient asexual propagation technique where the rootstock and scion from two plants are joined together to form a single plant. The production of grafted vegetable plants first began in Japan and Korea in the late 1920s with watermelons grafted onto pumpkin rootstocks. Now common in parts of Asia, Europe and the Middle East, grafting is used to improve traits like disease tolerance, abiotic stress resistance, and yield in important vegetable crops. Modern grafting methods include hole insertion grafting, tongue approach grafting and cleft grafting. Healing chambers are used to promote graft union formation and robotic grafting systems can produce hundreds of grafted plants per hour.
This document provides information on onion seed production including floral biology, methods, requirements, and standards. It discusses:
1) Onion flowers are protandrous, with pollen shed occurring before stigma receptivity. Cross-pollination by insects is needed for high seed yields.
2) The bulb-to-seed method is most common, involving selecting bulbs in one season for planting the next season for seed production. Care is taken to select true-to-type bulbs.
3) Seed production requires isolation distances between varieties, rogueing of off-types, and inspection to ensure varietal purity and high seed quality and yields.
This document discusses seed storability and viability prediction in important oilseed crops. It covers several key topics:
1) Factors that affect seed viability and storability during storage like moisture content, temperature, packaging material and storage structure. Lower moisture content and temperature helps extend seed life.
2) Methods for predicting seed viability like accelerated aging tests, mathematical models and nomographs that relate viability to moisture content and temperature over time.
3) Biochemical changes that occur during storage like lipid peroxidation and accumulation of free radicals that can damage cell membranes and organelles leading to loss of viability over time.
4) The importance of understanding these factors and developing strategies to optimize seed storage conditions and predict viability
The document discusses guidelines for releasing and notifying crop cultivars in India. It explains that releasing a cultivar makes it available for public cultivation and allows farmers to choose varieties, while notification regulates seed quality under the Seeds Act. The process involves variety evaluation through regional trials over multiple locations and years before the State and Central Variety Release Committees decide on release. Notified varieties can then be certified to ensure standard seed quality. Advantages of notification include compulsory certification for seed production and regulation of quality for seed sales. Examples of notified rice, wheat and black gram varieties in different states are also provided.
This document provides information about soybean breeding in three parts. It begins with an introduction to soybean including its scientific name, family, uses, protein and oil production. Next is a history of soybean originating in China and spreading to other countries. The document concludes with descriptions of soybean plants, their floral biology, selfing and crossing techniques, genetics of traits, and seed production and certification standards.
This document discusses sex expression in cucurbitaceous crops. It notes that cucurbits can have various sex forms ranging from hermaphroditic to monoecious to gynoecious. The major cucurbits like cucumber, bitter gourd, muskmelon, watermelon are typically monoecious. Gynoecious lines have been developed in some crops like cucumber and muskmelon through breeding. Sex determination is controlled by genes but can be modulated by environmental factors like temperature and photoperiod. Chemicals like silver nitrate and gibberellic acid can also induce staminate flowers.
This document discusses seed processing and storage. The objectives of seed processing are to improve quality by removing impurities, maintaining viability and vigor, making handling easier, and increasing value. Methods used for processing include drying, cleaning, grading, packaging, labeling, and treatment. Storage aims to preserve seeds under controlled conditions to prolong viability for long periods. Factors that affect seed longevity are seed type, quality, coat integrity, moisture content, and storage environment. Orthodox seeds can be stored long-term at low temperature and humidity while recalcitrant seeds require different storage methods.
Seed refers to a fertilized ovule containing an embryo that can develop into a new plant. Scientifically, seed is defined as a fertilized mature ovule covered by a seed coat. There are several types of propagating materials that are also considered seeds, including tubers, bulbs, rhizomes, roots, cuttings, and grafts. Seeds are classified into different categories based on their origin and intended use, including nucleus, breeder, foundation, and certified seeds, with each subsequent category representing a larger scale of multiplication while maintaining genetic and physical purity standards.
The document discusses the taxonomy, origin, evolution, and breeding of tomatoes. It notes that tomatoes originated in South America and were domesticated from wild cherry tomatoes. Important tomato breeding objectives include increasing yield, improving fruit quality and shelf life, and developing resistance to diseases and abiotic stresses. Breeding methods discussed are introduction, selection, hybridization, and biotechnology. The first genetically modified tomato, Flavr Savr, was developed to have increased shelf life but did not significantly improve firmness.
Seed processing is a vital part of ensuring high quality seed for end users. It includes cleaning, drying, treatment, packaging, and storage. The goals of seed processing are to reduce bulk, increase longevity by drying to a safe moisture level and treating with protectants, reduce variability in vigor, and improve uniformity in size and shape. The sequence of operations typically includes drying, receiving, pre-cleaning, conditioning, cleaning, separating, treating, weighing, bagging, and storage or shipping. Processing aims to separate inert materials and weed seeds from the seed lot while upgrading quality by eliminating damaged or low vigor seeds to obtain a high percentage of pure seed with maximum germination potential.
- Tomato (Lycopersicon esculentum) is the second most consumed vegetable in the world after potatoes. It is cultivated throughout temperate and tropical regions for its nutrient-rich fruit.
- Tomatoes originate from the Andean region of South America but were domesticated in Mexico. They were later spread worldwide by Europeans.
- Tomatoes have chasmogamous flowers that self-pollinate but can also cross-pollinate up to 47% depending on conditions. Manual pollination techniques include emasculation, pollen collection and application.
Traditional phenotypic methods and newer genotypic methods can both be used to identify bacteria. Phenotypic methods include gram staining, culturing, and analyzing biochemical characteristics and reactions. These methods have limitations as some bacteria cannot be cultured. Genotypic methods like MALDI-TOF, PCR, and microarrays identify bacteria based on their genetic material and can identify bacteria directly from clinical samples faster than phenotypic methods. A variety of biochemical tests are used as part of phenotypic identification to analyze carbohydrate metabolism, production of specific compounds, enzyme activity, and other characteristics.
This document discusses various methods for assessing genetic purity in plants, including morphological, chemical, and electrophoresis-based methods. Morphological methods involve examining seed or plant traits under magnification or in a grow-out test. Chemical methods analyze seed components like secondary metabolites and proteins. Electrophoresis separates proteins or DNA based on size and charge, allowing comparison of banding patterns between varieties. Together, these methods allow testing seed samples against a pure reference to validate their genetic purity or identify off-types.
This document describes a study on genetic diversity in soybean (Glycine max) using RAPD marker. Six soybean genotypes were collected and their DNA was extracted using the CTAB method. The DNA was quantified and found to range from 158-502 ng/μl. The objectives were to analyze genetic diversity among the genotypes using RAPD marker. The methodology involved DNA extraction, quantification, PCR amplification with RAPD primers, and resolving the amplified products through gel electrophoresis. The results showed good quality DNA was extracted. The study provides information on genetic variation that can aid in soybean breeding programs.
Genotoxicity_studies M pharmacy Pharmacology.pptxAyodhya Paradhe
This document discusses various genotoxicity studies and guidelines. It introduces the concept of genotoxicity and describes several important tests used in genotoxicity assessment, including the Ames test, in vitro mammalian cell micronucleus test, in vivo mammalian erythrocyte micronucleus test, in vitro mammalian chromosomal aberration test, and in vivo mammalian bone marrow chromosome aberration test. It provides details on the principles, procedures, and reporting of results for these key genotoxicity tests.
Detection of Genetically modified plants and Organic Seed production.NSStudents
The Presentation is prepared by the N.S Institution of science, Markapur.
It consists of a basic introduction related to Detection of Genetically modified plants and Organic Seed production.
This document summarizes different methods for testing genetically modified (GM) seed and trait purity, including DNA-based, protein-based, and bioassay methods. DNA-based methods include endpoint PCR, real-time PCR, and other technologies to detect the presence of GM DNA. Protein-based methods include lateral flow strip tests and enzyme-linked immunosorbent assays (ELISAs) to detect GM proteins. Bioassays involve growing seeds in controlled conditions and observing for trait expression. The document provides details on ELISA tests, lateral flow strips, electrophoresis, polymerase chain reaction (PCR), and considerations for calculating and expressing testing results.
molecular and biochemical for veritial identification SrijanLama3
This document discusses molecular and biochemical methods for varietal identification of seed. It describes electrophoresis, which separates proteins and enzymes extracted from seeds based on banding patterns that can differentiate varieties. PCR (polymerase chain reaction) amplifies specific DNA fragments, allowing detection and identification of genes through size and charge visualization via gel electrophoresis. Molecular markers like RFLPs and VNTRs also detect contamination and segregation by obtaining unique patterns for each variety. Biochemical methods like electrophoresis analysis of proteins and isozymes provide efficient, cost-effective cultivar identification and purity testing as seed storage proteins are independent of environmental fluctuations.
The production of haploid plants exploiting the totipotency of microspore.
Androgenesis is the in vitro development of haploid plants originating from totipotent pollen grains through a series of cell division and differentiation.
improved cultivation and metagenomics as new tools for bioprospecting in cold...Nicol Hormazabal
This document summarizes improved cultivation and metagenomic methods for bioprospecting in cold environments. Only a small percentage of microorganisms can typically be cultured using standard techniques, leaving much potential undiscovered. Improved cultivation methods aim to better mimic the natural environment, such as using diffusion chambers, hollow-fiber membrane chambers, and gel microdroplets to allow nutrient exchange while separating microbes. Metagenomic methods involve direct sequencing or functional screening of environmental DNA without cultivation. Both approaches can be combined, such as using DNA from enrichment cultures. Bioprospecting in cold environments faces additional challenges like low biomass and restricted access, requiring adaptation of methods.
Post genomic tools for genetic enhancement of germplasmVishu1234567
This document discusses how post-genomic tools like transcriptomics, proteomics, and metabolomics can be used for genetic enhancement of germplasm. It provides an introduction to each omics technique, examples of technologies used, and applications in understanding biological processes and identifying genes/proteins involved in traits. The conclusion states that omics expression analysis of germplasm will help characterize genome function and restore traits from wild varieties, aiding development of more sustainable crop varieties.
SOMA CLONAL VARIATION IN PERENNIAL HORTICULTURE CROP,.pptxPradeepti Sharma
1) Somaclonal variation refers to genetic variations that arise in plants regenerated from tissue culture. It can produce useful traits for plant breeding as well as undesirable variations.
2) Mechanisms of somaclonal variation include pre-existing genetic variations in somatic cells as well as new mutations generated during tissue culture due to stress conditions.
3) Somaclonal variation has been induced in many perennial horticulture crops, producing variants with traits like disease resistance and stress tolerance, though undesirable variations can also arise. Selection techniques can help recover useful variants.
OMICS is the comprehensive analysis of the biological system. The technologies which made a revolution such as Genomics, Transcriptomics, Proteomics, Metabolomics and Phenomics, in screening traits and develop novel improved organisms are mentioned here. The presentation gives a brief idea about various OMICS technology used in crop improvement, their steps, techniques used, applications, scope, advantages and disadvantages.
Plant Metabolism Studies: Options for Plant CultivationCovance
Regulators across the world are concerned with ensuring that any residues left in or on a crop after application of a plant protection product (PPP), present minimal risk to the health of humans and animals. To achieve this, regulators need information on the identity of the residues and the levels of residues remaining in or on a crop, in order to assess dietary risk and set maximum residue levels (MRLs). The testing approaches used are harmonized across most countries worldwide, focusing on the Organization for Economic Co-operation and Development (OECD) Test Guidelines (TGs) for pesticide residue chemistry. This e-book paper focuses on the laboratory-based plant cultivation methods that underlie the success of OECD crop metabolism studies, namely TG 501 and 502.
Presence of genetically modified organism genes in carica papaya, glycine max...valrivera
This document summarizes a study that aimed to detect the presence of genetically modified organism (GMO) genes in fruits from four plants: papaya, soybean, corn, and wheat. DNA was extracted from samples of each fruit and tested via polymerase chain reaction (PCR) and gel electrophoresis to detect two common GMO markers - the 35S promoter and NOS terminator. The results were inconclusive due to DNA degradation and possible human errors during experiments. As such, the study was unable to determine if the fruits contained GMO genes.
Presence of genetically modified organism genes in carica papaya, glycine max...Carlos Santos Perez
The document summarizes a study that aimed to detect the presence of genetically modified organism (GMO) genes in fruits from four plants - Carica papaya, Glycine max, Triticum spp., and Zea mays - using polymerase chain reaction (PCR) and gel electrophoresis. DNA was extracted from samples of each fruit and tested with PCR for the presence of common GMO markers. Gel electrophoresis revealed bands indicating the presence of plant DNA but unclear or missing bands for the GMO markers, likely due to DNA degradation. The results did not confirm or deny the hypothesis due to errors affecting the experiment.
Evaluation of protein and peptide formulations.pptxDivya Pushp
This document discusses stability testing and evaluation methods for protein formulations. Stability testing ensures products maintain specifications over shelf life under various storage conditions. Evaluations include bioassays to assess potency, which can be in vitro by monitoring cell responses to proteins or in vivo by monitoring animal pharmacological responses. Common evaluation methods are UV spectroscopy, Bradford assay, thermal analysis like DSC and TGA, and chromatography techniques like HPLC, ion exchange, and chromatofocusing.
The Ames test is a bacterial mutagenicity assay used to identify potential carcinogens. It involves exposing histidine-requiring strains of Salmonella typhimurium bacteria to test compounds or chemicals. If a compound is mutagenic, it will induce mutations in the bacteria that allow their growth in the absence of histidine. The Ames test is quick, inexpensive, and has identified many carcinogens and mutagens. However, its sensitivity is sometimes limited as bacteria do not fully replicate human metabolism.
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Advancements in seed testing technologies
1. Department of Seed Science and Technology
Chaudhary Sarwan Kumar Himachal Pradesh Krishi
Vishvavidyalaya , Palampur
Speaker - Jeenia Thalyari
Roll no - A-2018-30-070
3. Outline of the seminar
This presentation includes:
1. Introduction
2. History of seed testing
3. Concept of seed quality
4. Advances in genetic purity testing
5. Advances in physical purity testing
6. Advances in physiological quality testing
7. Advances in seed health testing
8. Conclusion
4. Introduction
“Subeejam Sukshetre Jayate Sampadayte"
• High quality seed is determined by its parent’s genetic makeup, physical
integrity , purity , its health and physiological quality.
• Farmers continue to adopt precision agricultural practices and depend
on the performance of every single seed put into the ground— the only
way to achieve that is by seed testing.
Every inch of the field counts, and seed testing plays a significant role.
5. Seed Technology encompasses:
1. Development of superior
varieties
2. Production
3. Evaluation
4. Processing
5. Storage
6. Testing
7. Certification/quality control
8. Marketing and distribution
9. Seed pathology
10. Seed entomology
11. Seed physiology
12. Seed ecology
6. Seed Testing
Seed testing is the art and science of evaluating seed quality for
agricultural purposes (Copeland and McDonald, 2001).
Seed tests tell us if a crop of seeds is worth collecting, if handling
procedures are correct, and how many potential seedlings are available
for regeneration.
• Seed quality in India is legally controlled by The Seeds Act, 1966.
7.
8. Seeds - basic, vital and crucial
input for crop production
Quality control of seeds depends
on the different seed testing
protocols which determine the
genuineness of the cultivar.
Evaluates the planting value and
the authenticity of the certified lot.
Assess the seed quality attributes
of the seed lots which have to be
offered for sale.
9. History of seed testing
• Seedquality test - seed germination and purity.
• First official seed testing station by Frederick Nobbe
(1869) in Tharandt (Germany)
• 1876- Lab established in USA - Connecticut
Agriculture experiment station.
• Now such labsestablishedin all countries. Professor F. Nobbe
10. o 1931:Germination, Purity,Sanitary conditions,
Genuineness of variety,weight determinations,
determination of moisturecontent.
o 1966:SeedHealth Methods
o 1968:Tetrazoliumtest
o 2001:Vigourtests
o 2004:Performancebased
methodsforspecifiedtrait testing
12. 1. Destructive
•Tests are carried out to understand the seeds performance
or behaviour under different loads.
• Example Tetrazolium test and vigour tests
13. 2. Slow
•Most of the methods require time for completion.
•Example germination test and vigour tests.
14. 3. Require pre-treatment
Viability tests like Tetrazolium test require pre-treatments
/ pre-conditioning of some seeds.
Example wheat seeds are kept between moist towels or
directly immersed in water for 18 hrs at 20°C.
Soaked wheat seeds Stained seeds
15. 4. Less accuracy
• Germination- “The emergence and development of the
seedling to a stage where the aspect of its essential
structures indicates whether or not it is able to develop
further into a satisfactory plant under favourable conditions in
the soil” (ISTA 2004)
• There are potentially many different interpretations of this
definition. Some variability within the interpretation of the rules
can lead to differences in the classification of a seed as normal
or abnormal.
17. Seed quality
• Trueness to variety
• Presence of inert matter
• Seeds of other crops
• Weed seeds
• Germination percentage
• Vigour
• Appearance
• Freedom from diseases
- aspects of seed quality.
Seed quality describes the potential performance of a seed lot.
18.
19. Genetic purity
Genetic purity is defined as true to type plants / seeds conforming to the
characteristics of the variety as described by the breeders.
It refers to the percentage of contamination by seeds or genetic material of
other varieties or species.
20. Methods to assess genetic purity
1. Morphological
2. Conventional grow out test
3. Chemical tests
4. Gel electrophoresis
5. Polymerase Chain reaction
6. Molecular markers(DNA)
(Basra, 2002)
21. Traditional approach to purity testing
Morphological traits
Seed morphology Examination of Seedlings
In field
In lab or greenhouse
22. CHEMOTAXONOMY
The use of biochemical methods to analyze various components of
seeds. Chemo taxonomists have recognized two groups of compounds
that are generally useful in classification of plant species:
1. Episemantic or secondary metabolites
(pigments or fatty acid etc.)
2. Semantides or sense carrying molecules
(Proteins, Nucleic Acids)
Methods of testing based on -
1. Analysis of secondary metabolites
2. Protein analysis
3. Nucleic acid analysis
23. These tests range from simple colour tests to complex
chromatographic separations of phenols, anthocyanin,
flavonoids and other compounds.
1. Phenol test- Wheat
2. Peroxidase test- Soybean
3. Potassium hydroxide – bleach test- Sorghum
4. Fluorescence test - Ryegrass
5. NaOH test- Wheat
6. Anthocyanin test- Rice, Soybean
1. Analysis of secondary metabolites
24. Proteins are the direct gene products, therefore the analysis of
seed, seedling proteins and enzymes is most successful and widely
used. Hence much attention has been focused on seed storage
proteins.
There are two primary methods:
1. Gel electrophoresis
2. Isoelectric focusing
2. Protein analysis
25. Gel electrophoresis
• The term gel electrophoresis refers to a technique used for
separation and analysis of DNA, RNA and proteins based
on their size and charge.
• The suffix phoresis means “migration” or “movement”,
while the prefix electro indicates the use of electricity as a
mean to separate molecules.
26. SDS-PAGE (Sodium dodecyl sulphate-
Polyacrylamide gel electrophoresis)
Denaturing and reducing sodium dodecyl
sulphate (SDS)-PAGE- widely used
electrophoresis technique
SDS-PAGE separates proteins primarily by
mass because the ionic detergent SDS
denatures and binds to proteins to make
them uniformly negatively charged.
Thus, when current is applied, all SDS-
bound proteins in a sample will migrate
through the gel toward the positively
charged electrode.
Proteins with less mass travel more quickly
through the gel than those with greater
mass - sieving effect of the gel matrix.
27.
28. Isoelectric focusing(IEF)
Isoelectric focusing (IEF)- separates molecules by differences in
their isoelectric point (pI).
Performed on proteins in a gel- takes advantage of the fact - overall charge on the
molecule of interest is a function of the pH of its surroundings.
A protein that is in a pH region below its isoelectric point (pI) will be positively
charged and so will migrate toward the cathode (negatively charged electrode).
As it migrates through a gradient of increasing pH, the protein's overall charge
will decrease until the protein reaches the pH region that corresponds to its pI.
At this point - no net charge and migration ceases.
As a result, the proteins become focused into sharp stationary bands with each
protein positioned at a point in the pH gradient corresponding to its pI.
29.
30. 3. Nucleic acid analysis
Polymerase chain reaction
Polymerase chain reaction (PCR) is an efficient and cost effective
molecular tool to copy or amplify small segments of DNA or RNA.
PCR was originally developed in 1983 by the American biochemist
Kary Mullis. He was awarded the Nobel Prize in Chemistry in 1993
for his pioneering work.
31. Procedure
1. Denaturation: In order to make a copy of the DNA, the strands are separated
using heat to break the bonds between complementary nucleotides.
Temp/time= 95°C for 30 seconds
2. Annealing: To add a primer that will serve to flag down the building crew –the
polymerase- so it knows where to start copying.
• For this PCR primer is added -a small piece of DNA that dictates which part of the
double helix will be copied.
Temp/time= 50°C for 30 seconds.
3. Extension: To add new complementary nucleotides. In PCR, Taq polymerase, a
thermostable polymerase is used instead of DNA polymerase.
• Taq polymerase adds the complementary nucleotides to the single-stranded DNA,
forming a new, duplicate double helix.
Temp/time= 72°C for 60 seconds
32.
33. DNA Markers
Grow out test
Based on the identification of
morphological characteristics at various
stages of plant growth.
Subject to influence by environmental
factors and is time-consuming.
DNA markers
Based on the genotype of the
hybrids eliminating environmental
variations .
Based on identification of genotype
specific profiles
34. SSR markers- Simple sequence repeats – Repeating Sequence of 2-5
nucleotides.
Microsatellite markers can be used for fingerprinting of hybrids,
assessing variation within parental lines and testing the genetic purity of
hybrid seed lot.
DNA markers systems, PCR based co-dominant SSRs (microsatellites)
are preferred for genotyping – reproducibility and abundance .
35. After isolation of DNA , total of 75 variable SSR primer pairs
distributed across the 10 chromosomes were used for PCR
amplification.
General procedure
The banding pattern through gel electrophoresis of all these hybrids
showed both the amplicons present in female as well as pollen
parent, thus confirming the genuine crossing and heterozygotic
condition of the hybrid.
The SSR markers identified had both female and male specific bands
and are useful in genetic purity testing.
1.
2.
3.
36.
37. Physical purity
Cleanliness of seed from other
crop seeds, weed seeds, inert
matter, diseased seed and insect
damaged seed.
Physically pure seeds should have
uniform size, shape, weight and
appearance.
Lack of this quality characters will
indirectly influence the field
establishment and planting value
of seed.
38. General method of purity separation
Place the sample on the purity work board after sieving / blowing
operations and separate the pure seeds.
After separation, we identify the pure seeds, each kind of weed
seeds, other crop seeds as to genus and species, inert matter -
names and number of each are recorded.
Seed blower Purity work board
39. 1. Image analysis
Image analysis is based on the extraction of data from a captured image for
characteristics like colour, size, shape of seed and seedlings and their subsequent
processing with the help of suitable computer software.
Speedy analysis, cost-effectiveness, automatic nature and user-friendly
environment for work are some important advantages .
Machine vision or computerized image analysis system is found to be very
convenient method for seed related studies as it is free from human errors.
40. 2.Ergovision inspection station
An improved ‘Microscopic Station’ that
incorporates advanced optical, ergonomic
and mechanical technologies to achieve fast
and accurate testing.
The main goals that were considered in
developing the new station were:
1.Accuracy, by providing the best
magnification, clarity and resolution for
each seed
2.Ergonomics, to reduce analyst’s fatigue
and discomfort
3.Productivity, to ensure fast and timely
results
- increased about 20-30% compared to the
traditional method.
41.
42. Physiological quality
It is the actual expression of seed in further generation /
multiplication.
• It comprises those intrinsic attributes of seeds which determine
their capacity to germinate and emerge rapidly and to produce a
uniform stand of vigorous plants under the range of field conditions
.
43. Germination and viability tests
1.Paper
2.Sand 3.Soil
a. Top paper
b. Between paper
c. Pleated paper
On Sand
Conventional methods- Germination
44. Conventional methods- Viability
1. Tetrazolium test- Common
2. Embryo excision test - Trees
3. Indoxyl acetate test - Soybean
4. Fast green test - Maize
5. Ferric chloride test - Legumes
46. 1. Micro-optrode technique
Micro-optrode technique (MOT) - invented by Porterfield and co
workers to measure seed viability in a quick and non-invasive
manner by measuring the oxygen influxes of intact seeds on the
cell surface.
There are two basic approaches :–
1. Long-term monitoring of oxygen consumption during seed
germination in an open system.
2. Detecting oxygen consumption by seeds by measuring the
decrease in oxygen concentration in a closed chamber within
10 seconds to screen one seed.
47. Procedure
Seeds soaked in solution (0.1 mM CaCl2, 0.1 mM KCl, pH 6.0) - three hours, and
then transferred to a new solution to detect oxygen fluxes.
A seed fixed on a Petri dish with a plastic colloidal cloth and immersed in
measuring solution.
The measuring point (embryo/embryonic axis) located by microscope.
Oxygen fluxes of seeds measured using micro-optrode calibrated in solution with
known oxygen concentrations .
The oxygen fluxes recorded every 10 seconds and measured for at least 5
minutes.
Final data of oxygen fluxes and the images acquired and recorded in real-time
using the imFlux software.
48.
49. 2. 3-dimensional X-ray imaging
High-resolution X-ray computed tomography (HRXCT) - important method
for non-destructive and non-invasive evaluation of seed internal structure.
How X-ray imaging works?
Three-dimensional X-ray imaging is based on the same principles as
conventional radiography.
1. X-rays produced by an X-ray generator are projected towards the object.
2. A certain amount of X-rays are absorbed by the object(seeds) depending
on its density and structural composition.
3. The X rays that pass through the object are captured behind the object
by a detector.
50. In computed tomography (CT) an X-ray source and its associated
detectors rotate around the object which itself moves around the
conical X-ray beam produced.
51. 3. Spectral imaging
Spectral imaging uses multiple bands across the electromagnetic spectrum.
An ordinary camera captures light across three wavelength bands in the visible spectrum- red,
green, and blue (RGB).
Spectral imaging encompasses a wide variety of techniques that go beyond RGB and may use
the infrared, the visible spectrum, the ultraviolet, X-rays, or some combination of the above.
It is possible to capture hundreds of wavelength bands for each pixel in an image.
52. Multispectral imaging captures a small number of spectral bands, typically three
to fifteen, through the use of varying filters and illumination.
A hyperspectral camera uses special hardware to capture hundreds of
wavelength bands for each pixel, which can be interpreted as a complete
spectrum.
53. Vigour tests
Seed vigour is an important quality parameter which needs to be
assessed to supplement germination and viability tests to gain insight
into the performance of a seed lot in the field or in storage.
ISTA congress in 1977 adopted the definition of seed vigour as " the
sum total of those properties of the seed which determine the level
of activity and performance of the seed or seed lot during
germination and seedling emergence".
54. Conventional methods
1. Growth tests
2.Conductivity test
3. Brick gravel test
4.Paper piercing test
5. Accelerated aging test
6.Cold test
Vigour tests
55. Seed Vigour Imaging System
Computerized image analysis, has made objective information
accessible in a relatively short period of time, with less human
interference.
Sako et al. (2001) developed an automated system for assessing the
vigour of lettuce seeds called the Seed Vigour Imaging System
(SVIS).
The process involves scanning the seedlings and then generating
vigour, growth and uniformity indexes by using certain softwares.
57. Chlorophyll fluorescence test
This method was first used by Jalink et al.(1998) on cabbage
seeds for the assessment of maturity and quality.
It is based on the non-destructive measurement of
chlorophyll- a in individual seeds.
During maturation for the majority of seed species the amount
of chlorophyll in the seed and seed coat decreases, whereas
the quality of the seeds increases.
Therefore, the quality of these seeds is inversely related to
the amount of chlorophyll in them.
58. This method uses the property of chlorophyll that it fluoresces
when it is excited at a certain wavelength.
Using this property and a combination of a red laser to
excite the chlorophyll and narrow bandwidth filter to filter out the
fluorescence, the chlorophyll in seeds can be determined.
59. Ethanol breath analyser
A fast and simple method to analyse
seed deterioration by measuring ethanol
production from partially imbibed seeds.
The test, uses a commercial breath
analyser, measures ethanol produced by
seeds in case of mitochondrial damage.
The analysis showed an inverse
correlation between ethanol production
and seed quality.
60. Seed health
Seed health is a measure of
freedom of seeds from
pathogens.
The presence or absence of
seed-borne pathogens can be
confirmed through the use of
seed health testing (Agrawal,
1995).
The term “seed health” includes
the incidence in the seed lot of
organisms like fungi, bacteria,
viruses, nematodes and insects.
61. Conventional methods of seed
health testing
Basic methods:
1. Direct Inspection
2. Washing Method
3. Incubation Methods
4. The whole Embryo Count Method
5. Blotter Methods
6. Agar Plate Test
7. Freezing Method
8. Growing Out test
9. Ordinary seedling symptom Test
Immunodiagnostic Methods:
1. Serology
2. Nucleic Acid based methods
Including PCR
3. Immunodiffusion Tests
4. Immunosorbent Electron
Microscopy (ISEM)
62. For insect pests
For plant pathogens
1. Alkali or glycerine method
4. New PCR techniques
3. ELISA
2. Latex flocculation test
1. Double diffusion test
Technological advancements
63. 1. Double diffusion test
Seed or part (embryo)
of it is ground to a fine
powder.
Triturate is transferred
to a well cut in a
diffusion media (agar
gel).
Antiserum specific
for a suspect virus
in a seed is placed
in separate well.
In time the virus particles
(antigen) and antibody
molecules diffuse towards
one another.
Since diffusion is in two
direction it is called
Double diffusion.
Diffusion confirms
the presence of
virus.
64. 2. Latex agglutination test
Grinded seed extracts are
placed in a pipette
Specified quantity of tagged
latex is added
Pipette is then oscillated
for about 15 minutes
Observed under
microscope
When pathogens are present in the
sample the latex suspension
becomes flocculated .
1.
2.
3.
4.
5.
65. 3. Enzyme Linked Immunosorbent
Assay(ELISA)
ELISA (Enzyme-linked immunosorbent assay) is a plate-based assay
technique designed for detecting and quantifying peptides, proteins,
antibodies and hormones.
In ELISA, an antigen is immobilized to a solid surface and then complexed
with an antibody that is linked to an enzyme.
66.
67. The authors mentioned various common methods of detecting seed
borne pathogens along with some recent methods :
1. Bio-PCR
2. IMS-PCR
3. Real time PCR
68. 1. Bio-PCR
• Bio-PCR is culturing of bacterial cells to the detectable limit
prior to PCR. This in turn increases sensitivity and allow early
detection of bacterial pathogens.
• Different plant pathogens Ex. Pseudomonas syringae pv.
Phaseolicola, Acidovorax avenae spp. Avenae, Xanthomonas
oryzae pv. Oryzae can be detected by Bio-PCR method.
70. 3. Real time PCR
This method consists of coupling DNA amplification with fluorescence
substances which can be easily measured, giving an indirect measurement
of DNA amplification.
Procedure-
1. The PCR is prepared as usual , and the reporter probe is added.
2. As the reaction commences, during the annealing stage of the PCR both
probe and primers anneal to the DNA target.
3. Polymerization of a new DNA strand is initiated from the primers, and
once the polymerase reaches the probe, its 5'-3'-exonuclease degrades
the probe, physically separating the fluorescent reporter, resulting in
an increase in fluorescence.
4. Fluorescence is detected and measured in a real-time PCR machine.
73. 1. Seeds are taken in a beaker (100 mI capacity) and then 10% Sodium hydroxide
solution is poured in it until the seeds are submerged.
2. The seeds are boiled in Sodium hydroxide solution for 10 minutes.
3. After decanting solution, translucent seeds are washed with water and then
examined with the help of magnifying glass.
4. Seeds with visible internal infestation are separated, cut open to confirm the
presence of insect and counted to report the result in percentage (by number)
74. Conclusion
The main feature of the mentioned technologies is that most of
them are multi- purpose and therefore can be used for carrying
out more than one component of seed quality.
Spectral
imaging
Viability
Damage
detection
Seed Health
PCR
Genetic
purity
Seed health
3-D X-ray
imaging
Physical
purity
Viability
Seed health
75. Safeguarding seed quality is the first step towards a satisfying yield.
It is estimated that the direct contribution of quality seed alone to the
total production is about 15-20% depending upon the crop species.
The determination of seed quality by seed testing through advanced
technologies is the most important measure for better crop production.