A transplastomic plant is a genetically modified plant in which the new genes have not been inserted in the nuclear DNA but in the DNA of the chloroplasts.
Roughly based on Chapter 11 Biotechnology: Principles and Processes and Chapter 12 Biotechnology and its Applications of Class 12 NCERT for final brush-up before the exams
Topics included - Introduction; explanation; examples like blue white screening method, antibiotic resistance; Extra information regarding - detection of oncogene in vertebrates and role of sleeping beauty; Merits and demerits of insertional inactivation.
A transplastomic plant is a genetically modified plant in which the new genes have not been inserted in the nuclear DNA but in the DNA of the chloroplasts.
Roughly based on Chapter 11 Biotechnology: Principles and Processes and Chapter 12 Biotechnology and its Applications of Class 12 NCERT for final brush-up before the exams
Topics included - Introduction; explanation; examples like blue white screening method, antibiotic resistance; Extra information regarding - detection of oncogene in vertebrates and role of sleeping beauty; Merits and demerits of insertional inactivation.
The ultimate objective of modern plant breeding is to improve a top variety in one single additional character in a predictable and precise manner without disturbing the rest of the genome. Today this is being realised through examples of successful transfer of specific traits into higher plants by gene transfer.
Techniques that open up to the plant breeder the possibility of transferring in a planned manner characters from one organism to another have been developed in microbial genetics. It should be stressed right at the outset that the expression “gene” has different meanings in agriculture and in molecular biology.
Gene Transfer Methods:
The gene transfer techniques in plant genetic transformation are broadly grouped into two categories:
I. Vector-mediated gene transfer
II. Direct or vector less DNA transfer
It highlights the various methods of gene transfer in plants, characterization of plants by PCR and qRTPCR. Different types of PCR and Real time PCR have been described
This technique helps in maintaining plant tissues for prolonged periods on an artificial culture/basal medium. Small group of cells is isolated and inoculated on a culture medium comprising carbohydrate source, vitamins, minerals and defined nitrogenous source(s) under sterilized lab conditions.
It is possible to cultivate any plant species using variable explants e.g. stem segment, leaf segment, pedicel, petiole, anther, pollen, microspores, petiole, cotyledons, endoperm, etc. Callus derived from a single initial group of cells can be sub-cultured, multiplied and in some instances induced to reinitiate differentiation of roots and shoots and thus form plantlets (Fig. 30-1). The latter are hardenend and then transferred to the soil.
The ultimate objective of modern plant breeding is to improve a top variety in one single additional character in a predictable and precise manner without disturbing the rest of the genome. Today this is being realised through examples of successful transfer of specific traits into higher plants by gene transfer.
Techniques that open up to the plant breeder the possibility of transferring in a planned manner characters from one organism to another have been developed in microbial genetics. It should be stressed right at the outset that the expression “gene” has different meanings in agriculture and in molecular biology.
Gene Transfer Methods:
The gene transfer techniques in plant genetic transformation are broadly grouped into two categories:
I. Vector-mediated gene transfer
II. Direct or vector less DNA transfer
It highlights the various methods of gene transfer in plants, characterization of plants by PCR and qRTPCR. Different types of PCR and Real time PCR have been described
This technique helps in maintaining plant tissues for prolonged periods on an artificial culture/basal medium. Small group of cells is isolated and inoculated on a culture medium comprising carbohydrate source, vitamins, minerals and defined nitrogenous source(s) under sterilized lab conditions.
It is possible to cultivate any plant species using variable explants e.g. stem segment, leaf segment, pedicel, petiole, anther, pollen, microspores, petiole, cotyledons, endoperm, etc. Callus derived from a single initial group of cells can be sub-cultured, multiplied and in some instances induced to reinitiate differentiation of roots and shoots and thus form plantlets (Fig. 30-1). The latter are hardenend and then transferred to the soil.
Transgenes may be used to produce GMS which is dominant to fertility.
In these cases it is essential to develop effective fertility restoration systems for hybrid seed production.
An effective restoration system is available in at least one case, Barnase/Barstar system
Recombinant DNA techniques have made it possible to engineer new systems of male sterility by disturbing any or number of developmental steps specifically required for the production of functional pollen within the microspore or for the development of any somatic tissues .
Efficient, quick and tissue culture independent system for crop plants improvement useful for those plants that lack tissue culture and regeneration system.
Two most common Agrobacterium mediated in-planta methods such as floral dip and vacuum infiltration have been successfully used by many researchers in both dicot and monocot plants.
Main advantages of in-planta transformation are to produce large number of transgenic plants and accumulation of high concentration of total soluble protein in short time.
The presentation describes the advantages of plastid transformation over 'conventional' nuclear transformation, hurdles to plastid transformation, its advantages. The presentation also covers some successful plastid engineering and its potential.
Agrobacterium and other methods of plant transformation including gene gun, i...PABOLU TEJASREE
The process of transfer, integration and expression of transgene in the host cells is known as genetic transformation. A foreign gene (transgene) encoding the trait must be incorporated into plant cells, along with a "cassette" of extra genetic material to add a desirable trait to a crop. The cassette includes a sequence of DNA called a "promoter", which determines where and when the foreign gene is expressed in the host, and a "marker gene" which allows breeders to determine by screening or selection which plants contain the inserted gene. For example, marker genes may make plants resistant to antibiotics not used routinely (e.g., agrimycin, kanamycin) or tolerant of some herbicides.
Assessment of genetic fidelity of in vitro propagated clones of Celastrus pan...iosrjce
Celastrus paniculatus Willd belonging to the family Celastaceae is an endangered Indian medicinal
plant having high pharmaceutical application. The objective of the present investigation was to assess the the
clonal fidelity of in vitro propagated clones of Celastrus paniculatus with the field grown mother plant to
confirm their true to type nature. Micropropagation is an alternative method for the large scale production of
endangered medicinal plants. The genetic stability of in vitro raised clones of celastrus paniculatus were
assessed by using RAPD analysis. Genomic DNA was isolated from healthy and fresh leaves of both mother
plant and in vitro raised plants of Celastrus paniculatus by using CTAB method. Based on the reproducibility of
the primers, 15 RAPD primers were selected for the present investigation. The selected primers gave rise to a
total of 75 scorable bands with an average of 5.1 bands ranging from 300-2700 bp. The number of bands varied
from three (OPQ-07, OPA-13) to seven (OPC-20, OPN-16). Randomly selected 10 micropropagated plants
from each culture period was used. Amplification pattern was electrophoresed in 1.5% TBE, revealing that all
the bands produced by micropropagated plants were monomorphic and similar to that of the field grown plant.
No polymorphism was detected by RAPD analysis.
To study of the genetic variations among the Azospirillum lipoferu isolates u...ijsrd.com
Among free-living microorganisms, which can be practically used in agriculture, bacteria from the Azospirillum genus as well as other endophytes are nowadays thought of as the most active component of associative dinitrogen fixation. The investigation was carried out to study the characterization of Azospirillum lipoferu found in the soils of the ten agro-climatic zones which Karnataka, is classified. By using RAPD markers, 75 bands were scored out of which 78.6 % were found to be polymorphic. Statistical analysis of RAPD data enabled the classification of 10 Azospirillum isolates into two major groups. . In this, the cluster analysis based on 75 RAPD bands revealed that the ten A. lipoferu isolates examined clustered at a linkage distance of about 40 units on the dendrogram. There was no correlation between RAPD and geographical origin of isolates.
Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
A Strategic Approach: GenAI in EducationPeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
4. Ornamental floriculture is becoming an
important industry .
Ornamentals include a large variety of
crop plants
Cut flowers,
Bulbs and corms,
Foliage and Flowering pot plants.
All the present day ornamental varieties
and novelties are as a result of extensive
hybridization, induced mutation and
selection .
5.
6. Genetic engineering:The technology of
preparing recombinant DNA in vitro by
cutting up DNA molecules and splicing
together fragments from more than one
organism.
Genetic engineering is a laboratory
technique for gene manipulation.
Genetic engineering brings about novel
combination of genes by using
recombinent DNA technology which is not
7. Genetic engineering of plants is much
easier than animals.
there is natural transformation system for
plants(Agrobacterium).
plant tissue can redifferentiate.
plant transformation and regeneration
are relatively easy for a variety of plants.
Agrobacterium tumefaciens can infect
wounded plant tissue, transferring a
large plasmid, the Ti plasmid, to the
plant cell.
8. Important methods in
recombinant DNA technology are
Isolation of desired gene Insertion
of isolated gene into a suitable
vector Introduction of recombinant
vector in to host Selection of
transformed host cells
(A.C.Dutta 2005)
9. Digestion of the cell wall by enzymatic
action, dissolution of the biological
membranes by detergent losses,
centrifugation to isolate pure DNA.
DNA cut into no. of fragments by restriction
endonulcleases “molecular scissors” with
sticky ends.
11. Most widely used
More economical
More efficient
Agrobacterium mediated
gene transfer
Particle bombardment
or
micro projectile .
Direct DNA delivery by
PEG .
Electroporation .
Microinjection .Chandler and Brugliera, 2011
12.
13. 1-2 µm of tungsten or gold particles
(microprojectiles)coated with DNA to be used for
transformation are accelerated to velocities using
pressurized Helium gas
14. DNA solution is injected directly inside the cell
using capillary glass micropipetts .
24. Plant
material
Surface
sterilization
Morphogen
esis
Induction
Medium
(MIM)
Rooting
Acclimatization
•After 2.5 - 3 months
in culture, calli
showing well
developed
morphogenetic
structures (shoots in
the case of P. zonale
and somatic embryos
in P. peltatum) were
transferred to a
selective Elongation
Medium .
• Elongation
Medium (EM: MS
basal medium and
Shahin vitamins,
supplemented with
50 mg l-1
kanamycin)
• All explants
were subculture
every 2 weeks
onto the same
fresh medium until
shoots were long
enough to be
separated ..
Elongation
Medium
26. Plant
material
Surface
sterilization
Morphogen
esis
Induction
Medium
(MIM)
Elongation
Medium
Rooting
•and acclimatized
in growth
chambers under
(16-h light/8-h
dark photoperiod)
and then
transferred to a
greenhouse until
they flowered..
• Regenerated
plantlets with
welldeveloped
roots were
transferred to
plastic pots
containing peat
moss and perlite
(3:1).
Acclimatization
Transformation efficiency was
estimated
as the number of independent
transformation
events (one transgenic plant per
explant) in relation to the total
number of inoculated explants.
27. Cytokinins have been implicated in several aspects of
plant development, including plant senescence [15-
20],
and are thought to be synthesized mainly in the roots
and transported to the shoots via the xylem.
Overexpression of the ipt gene in transgenic plants led
to elevated foliar cytokinin concentrations and delayed
leaf senescence, but high cytokinin levels have been
reported to be detrimental to growth and fertility [26
30].
To circumvent these effects :
Specificgene promoter (pSAG12 )
28. Promoter which
induces transcription
in male reproductive
specifically
Gene which disrupts
normal function of cell
Agrobacterium-
mediated
transformation
regeneration
male-sterile
plant
29. (A portmanteau of "BActerial" "RiboNucleASE")
is a bacterial protein that consists of 110 amino acids
and has ribonuclease activity.
It is synthesized and secreted by the bacterium
Bacillus amyloliquefaciens, but is lethal to the cell
when expressed without its inhibitor barstar .
The inhibitor binds to and occludes the ribonuclease
active site, preventing barnase from damaging the
cell's RNA
30.
31.
32. • LBA4404 cells were electroporated
to carry different plasmids a pBIN19 binary vector .
34. Bacteria were grown at 28°C on solid LB
plates supplemented with 40 mg l-1
rifampicin and 100 mg l-1 kanamycin
Single colony was used to inoculate 25 ml of LB
liquid medium with the same antibiotics ,
maintained at 28°C and 200 rpm for 24 h
Inoculate a liquid MS medium supplemented
0.2 mM acetosyringone dissolved in 70%
ethanol (sterilized by filtration), which was
cultured at 28°C for 12 h.
Inoculation of explants was conducted in
bacterial culture
35.
36.
37.
38.
39. Transformed explants were examined periodically for
gfp expression under a fluorescence stereomicroscope
(Leica MZ FLIII) .
40.
41.
42.
43. Identification of the ipt transgene (460 bp fragment) by PCR in different P. zonale pSAG12::ipt
transgenic plants. C + (positive control: pVDH393-pSAG12::ipt) and TI (negative control).
44. Identification of the barnase-barstar transgene (544 bp fragment) by PCR in different P.zonale male
sterile plants. C + (positive control: pBI101-PsEND1::barnase-barstar) and TI (negative control).
45. Realtime RT-PCR analysis of pSAG12::ipt transcript levels in detached leaves from the transgenic lines
3.4, 3.9, 4.3 and 4.12. Each sample’s expression level relative to Pelargonium x hortorum PhACTIN7 is
the mean of three biological repeats. C: control WT leaves.
46. Measurements were taken in the greenhouse on transgenic plants and
WT control plants :
Plant height (distance from soil line to top of the tallest growing point),
leaf length and width (average measurements from five fully expanded leaves),
leaf petiole length,
internodal length
Number of inflorescences per plant were evaluated.
Morphological measurements were taken over the course of several days on
each plant as its first five flowers reached anthesis .
Means differing significantly were compared at a 5% probability level.
Data variability was expressed as the mean ± SE.
47.
48.
49.
50.
51.
52. (a), 6 (b), 8 (c), 17 (d), 22 (e), 24 (f), 27 (g) and 34 (h) days of incubation in darkness.
53. Analysis of leaf senescence was conducted by extraction of
chlorophyll in detached leaves incubated in darkness from WT
control and pSAG12::ipt plants respectively.
Using a porcelain mortar cooled with liquid nitrogen,
samples were crushed to a fine powder. In 10 ml centrifuge tubes
the samples were mixed with 100 mg of MgCO3 and 5 ml of
100% (v/v) acetone. Bleached leaf material was removed by
centrifugation (5 min; 2,000 g) and 1 ml aliquots of supernatants
transferred to new tubes. Chlorophyll (a + b) content of extracts
was determined spectrophotometrically [53].
54. (i) Mean concentration (±SE) of chlorophyll a + b (mg/g fresh weight) from detached
leaves of control (WT) and pSAG12::ipt (TRG) plants at 0, 6 and 8 days of incubation
in darkness .
55. (j) Senescence delay of detached leaves from pSAG12::ipt plants. Fresh weight changes in detached
leaves of WT P. zonale and a transgenic line carrying the pSAG12::ipt chimaeric gene over the
time course analyzed. Data are the means of sixteen leaves ± SE. Bars: 1 cm.
56.
57. The chimaeric pSAG12::ipt construct useful in
Pelargonium spp. to delay the senescence process
and to produce long-lived plants, which could
have commercial interest.
Transgenic pSAG12::ipt plants showed
delayed leaf senescence, increased
branching and reduced internodal length
as compared to non-transformed plants.
Transgenic pSAG12::ipt plants showed a more
compact architecture than the WT.
58. Expression of the barnase gene under control of
PsEND1 promoter caused specific ablation of the
tissues, necrotic at early stages of anther
development.
No pollen grains were observed in the
ablated anthers from the male-sterile
plants, indicating that barnase effectively
destroys specific cell lines that form the
structural tissues of the anther , preventing
pollen development. .
59. The use of engineered male sterility
would be especially useful to
eliminate pollen allergens and to
produce environmentally friendly
transgenic plants carrying new
traits by preventing gene flow
between the genetically modified
ornamentals and related plant
species.
60. 50. Rogers SO, Bendich AJ: Extraction of total cellular DNA from plants, algae and fungi. Plant Mol Biol
Manual 1994, D1:1–8.
51. He J, Gray J, Leisner S: A Pelargonium ARGONAUTE4gene shows organspecific express
53. Lichtenthaler HK: Chlorophylls and carotenoids: Pigments of photosynthetic biomenbranes. Met Enzymol
54. Elliot AR, Campbell JA, Dugdale B, Brettell RIS, Grof CPL: Green fluorescent protein facilitates rapid in
vivo detection of genetically transformed plant cells. Plant Cell Rep 1999, 18:707–714.
55. Escobar MA, Park JI, Polito VS, Leslie CA, Uratsu SL, Mc Granahan GH, Dandekar AM: Using GFP as a
scorable marker in walnut somatic embryo transformation. Ann Bot 2000, 85(6):831–835.
56. Ghorbel R, Juárez J, Navarro L, Peña L: Green fluorescent protein as a screenable marker to increase the
efficiency of generating transgenic woody fruit plants. Theor Appl Genet 1999, 99:350–358.
57. Pérez-Clemente RM, Pérez A, García L, Beltrán JP, Cañas LA: Transformation and regeneration of peach
plants (Prunus persica L.) from embryo sections using the green fluorescent protein (GFP) as a vital marker. Mol
58. Rakosy-Tican E, Aurori CM, Dijkstra C, Thieme R, Aurori A, Davey MR: The usefulness of the gfp reporter
gene for monitoring Agrobacteriummediated transformation of potato dihaploid and tetraploid genotypes.
59. Yancheva SD, Shlizerman LA, Golubowicz S, Yabloviz Z, Perl A, Hanania U, Flaishman MA: The use of
green fluorescent protein (GFP) improves Agrobacterium-mediated transformation of ‘Spadona’ pear (Pyrus
60. Baranski B, Klocke E, Schumann G: Green fluorescent protein as an efficient selection marker for
Agrobacterium rhizogenes mediated carrot transformation. Plant Cell Rep 2006, 25:190