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International Journal of Trend in Scientific Research and Development (IJTSRD)
Volume 6 Issue 4, May-June 2022 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470
@ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1049
Soil Mycofloral Diversity of Wheat Crop Field of Kota Region
Amita Mali, Dr. Neerja Shrivastava
Department of Botany, Government College Kota, Kota, Rajasthan, India
ABSTRACT
Soil mycoflora are ecologically important for most crops because
they benefit plant growth and survival. The present study was
conducted with an objective to assess soil fungi diversity in field soil
in Kota, Rajasthan, and their role in growth and development of
wheat crop. The soil samples from different sites; where wheat crop
is grown in Kota region were collected, and fungal species were
identified, alongwith spore density calculation. The overall spore
density in wheat crop field in Kota region showed that the soil
properties were better due to mycoflora or fungal species diversity as
compared to field region without fungal diversity. The fungi
conservation is the basic aspect of this investigation. Mycoflora
provides fertility to soil and improves its quality increasing the yield
of wheat (Triticum aestivum) crop.
KEYWORDS: mycoflora, soil, diversity, Kota, wheat, crop, spore,
species, fungi, growth, yield
How to cite this paper: Amita Mali | Dr.
Neerja Shrivastava "Soil Mycofloral
Diversity of Wheat Crop Field of Kota
Region" Published
in International
Journal of Trend in
Scientific Research
and Development
(ijtsrd), ISSN: 2456-
6470, Volume-6 |
Issue-4, June 2022,
pp.1049-1054, URL:
www.ijtsrd.com/papers/ijtsrd50266.pdf
Copyright © 2022 by author(s) and
International Journal of Trend in
Scientific Research and Development
Journal. This is an
Open Access article
distributed under the
terms of the Creative Commons
Attribution License (CC BY 4.0)
(http://creativecommons.org/licenses/by/4.0)
INTRODUCTION
Research was conducted on isolation and
identification of soil Mycoflora of wheat fields in
Kota region. The soil samples used for the study were
collected from different field sites located in Kota
where only wheat (Triticum aestivum)crop was
grown.
IJTSRD50266
International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1050
The soil sample inoculation was carried was carried out using potato dextrose agar media. A total number of five
fungi were isolated Aspergillus niger, Aspergillus fumigatus, Aspergillus oryzae, Rhizopus oryzae and Rhizopus
stolonifer. Aspergillus fumigatus and Aspergillus niger had the highest percentage frequency of occurrences of
33.33%, Rhizopus oryzae and Rhizopus stolonifer had the lowest percentage frequency of occurrence of 6.67%
respectively. Conclusively the amount of work already accomplished in the investigating the micro flora of the
soil it must be admitted that no one has yet been able to give a clear picture of all life in the soil and of all
interaction of different groups of living things. Developing novel methodologies to study soil ecology under the
natural condition in wheat fields is needed and will required collaboration between plant biologists, ecologists,
and soil scientist to develop the a system where biochemical and molecular biology studies could be performed
on site. It has been estimated that one gram of surface soil contain 50,000 to a million fungi, the loss of organic
material from root provides the energy for the development of active fungal population in the Rhizosphere
around the root of wheat crop.
Wheat (Triticum aestivum) crop field in Kota
Methodology
The soil samples from wheat fields of Kota region were introduced into a test tube containing 9ml of sterile
distilled water. Suspension prepared was labeled and vigorously shaken. Serial dilution was prepared by
pippeting 1ml from stock suspension and inoculates in to 9ml of sterile distilled water in a labeled test tubes and
shaking vigorously to obtain 10-9
dilution 1ml was further pipette from the first dilution and introduced in to
another labeled test tube to obtain 10-1
dilution. They were repeated done to obtain up to 10-9
dilution
0.1ml each from 10-4
and 10-5
dilutions were inoculated in PDA plates and swabbed to ensure even distribution
of inoculums in the medium. Also 0.5ml each was taken directly from the stock suspension and inoculated into
PDA agar plate and swabbed as above. All plate were labeled and incubated at room temperatures for 5 to 6
days. After incubation the fungal colonies will be developed
Fully developed colonies with different cultural characteristic were subculture using sterile inoculating needles
in to freshly prepared PDA plates. Each colony was considered to have originated from one fungal propagules
Sub cultured plates were incubated at room temperatures for 3–5 days to obtain pure cultures of the various
fungal types.
International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1051
Fungal colonies in petriplates
Placing a drop of fungal stain using mounting needle on the clean slide and carefully teased (spread). A drop of
lacto phenol cotton blue was added on fungal stain and cover slip was placed on top. Then, examine under
microscope using x40 objectives The fungi were identified by way of microscopic method
Pure fungal cultures in test tubes
Discussion and Results
The fungal species isolated and identified are described in table below:-
Table-1: Morphological and Cultural Characteristic of the Isolated Fungi
Fungal Isolate Colony Texture
Aspergillus niger Colonies consisting of a compact white yellowish with dense of dark brown to black.
Aspergillus
fumigatus
Colonies consisting of a dense felt of dark green conidiophores intermixed with aerial
hyphae
Rhizopus
stolonifer
Colonies whitish becoming-greenish due to the brownish sporangiosphores and
brown black sporangia.
Rhizopus oryzae
Colony whitish becoming-gray with age, about 10mm high. Stolons smooth or
slightly rough almost colorless almost too yellowish brown.
Aspergillus
oryzae
Colonies consisting of a felt of long conidiophores often intermixed with aerial
mycelium. Pale greenish yellow, later becoming light to dull brown.
International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1052
Aspergillus niger
Hyphae are septate and hyaline. Conidial heads are
radiate initially, splitting into columns at maturity.
The species is biseriate (vesicles produces sterile cells
known as metulae that support the conidiogenous
phialides). Conidiophores are long (400-3000 µm),
smooth, and hyaline, becoming darker at the apex and
terminating in a globose vesicle (30-75 µm in
diameter). Metulae and phialides cover the entire
vesicle. Conidia are brown to black, very rough,
globose, and measure 4-5 µm in diameter
Aspergillus fumigatus
Aspergillus colonies are downy to powdery in texture.
The surface color may vary depending on the species.
The reverse is uncolored to pale yellow in most of the
isolates. Aspergillus fumigatus is a thermotolerant
fungus and grows well at temperatures over 40°C.
This property is unique to Aspergillus fumigatus
among the Aspergillus species. Aspergillus fumigatus
can grow at a temperature range of 20 to 50 °C.
Aspergillus oryzae
Other microscopic structures include sclerotia,
cleistothecia, aleuriconidia, and Hulle cells. These
structures are of key importance in identification of
some Aspergillus species. Cleistothecium is a round,
closed structure enclosing the asci which carry the
ascospores. The asci are spread to the surrounding
when the cleistothecium bursts. Cleistothecium is
produced during the sexual reproduction stage of
some Aspergillus species. Aleuriconidium is a type of
conidium produced by lysis of the cell that supports
it. The base is usually truncate and carries remnants
of the lysed supporting cell. These remnants form
annular frills at its base. Hulle cell is a large sterile
cell bearing a small lumen. Similar to cleistothecium,
it is associated with the sexual stage of some
Aspergillus species.
Rhizopus oryzae
Some morphological features, such as the length of
rhizoids and sporangiophores, the diameter of
sporangia, the shape of columellae, and the size,
shape and surface texture of sporangiospores aid in
differentiation of Rhizopus species from each other.
Rhizopus stolonifer
Nonseptate or sparsely septate broad hyphae (6-15
µm in diameter), sporangiophores, rhizoids (root-like
hyphae), sporangia, and sporangiospores are
visualized. Sporangiophores are brown in color and
usually unbranched. They can be solitary or form
clusters. Rhizoids are located at the point where the
stolons and sporangiophores meet. Sporangia (40-350
µm in diameter) are located at the tip of the
sporangiophores. They are round with flattened bases.
International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470
@ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1053
Apophysis is absent or rarely apparent and columellae
are hemispherical. Sporangiospores (4-11 µm in
diameter) are unicellular, round to ovoid in shape,
hyaline to brown in color, and smooth or striated in
texture
Table 2 Frequency of Occurrences of the
Isolated Fungi
Fungal Isolate Number of Occurrences
Aspergillus niger 5
Aspergillus fumigatus 5
Aspergillus oryzae 3
Rhizopus oryzae 1
Rhizopus stolonifer 1
TOTAL 15
Table 3 Percentage Frequency of Occurrences of
the Isolated Fungi
Fungal Isolate
Percentage of
occurrences (%)
Aspergillus fumigatus 33.33%
Aspergillus niger 33.33%
Aspergillus oryzae 20.00%
Rhizopus oryzae 6.67%
Rhizopus stolonifer 6.67%
Total Percentage 100%
The result obtained in this research indicated that
Aspergillus fumigatus and Aspergillus niger has the
higher percentages and high number of occurrences.
The highest number of fungal species isolated from
the rhizosphere of wheat crop is not surprising, due to
the production of substrate by growing root in the
form of root exudates containing amino, sugar,
organic acid, nucleotide and other substrate. Thus,
this lead to their proximity to the roots and
subsequent proliferation and multiplication of the
mycoflora in the soil region. Also high rate of
microbial decomposition of both organic and
sloughed off tissue (wheat) is yet another factor of
their abundance compared with the field soil where
such activities are minimal.
The fungal flora acts as universal agent of decay,
from which new life continually arises and is
nourished, the fungi makes a unique contribution to
the maintenance of soil fertility. Apart from the
saprophytic fungi which act as scavengers, a number
of important plant parasitic fungi live in the soil and
attack economically, important plants through their
roots or at the ground level. The obligate saprophytic
fungi isolated were genera of Aspergillus which affect
the wheat crop only when they are kept during
storage period. Also they add to the total biomass of
fungi in the soil and advantageous in the
decomposition of organic matter which enhances soil
fertility. It was also noted that, the intense microbial
activity in the soil (i.e. from both harmful and
beneficial microbes) can be especially important
because it may result in suppression or even
elimination of photogenic microorganism. Rhizopus
oryzae affects the wheat plants rottening it.
Conclusion
The result of this study indicated that five fungal
isolate where isolated which include the Aspergillus
niger, Aspergillus fumigatus, Aspergillus oryzae,
Rhizopus oryzae and Rhizopus stolonifer, which
indicate that out of these fungal isolate Aspergillus
niger and Aspergillus fumigatus has the higher
percentage frequency of occurrence of 33.33%,
followed by Aspergillus oryzae which has 20.00%,
while the Rhizopus oryzae and Rhizopus stolonifer
has the least percentage frequency of occurrence of
6.67% respectively. Thus it can be understood that
fungi are also necessary evils. They can be bound in
rhizosphere of crop and create fertility in field and
can also simultaneously be parasitic to wheat plants.
Hence useful fungi should be added to the field and
conservation should be done whereas harmful
parasitic fungi destroying crops should be treated
using antifungals of specific types. Mycoflora in
fields of wheat or any crop thus in general decay and
make soil fertile making growth and development of
plants more effective and in high frequency of fungal
species the rate of growth of crops also increases.
Hence positive utilities of mycoflora in soil makes
farmers to add good saprophytic decaying species in
fields in Kota region
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Soil Mycofloral Diversity of Wheat Crop Field of Kota Region

  • 1. International Journal of Trend in Scientific Research and Development (IJTSRD) Volume 6 Issue 4, May-June 2022 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470 @ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1049 Soil Mycofloral Diversity of Wheat Crop Field of Kota Region Amita Mali, Dr. Neerja Shrivastava Department of Botany, Government College Kota, Kota, Rajasthan, India ABSTRACT Soil mycoflora are ecologically important for most crops because they benefit plant growth and survival. The present study was conducted with an objective to assess soil fungi diversity in field soil in Kota, Rajasthan, and their role in growth and development of wheat crop. The soil samples from different sites; where wheat crop is grown in Kota region were collected, and fungal species were identified, alongwith spore density calculation. The overall spore density in wheat crop field in Kota region showed that the soil properties were better due to mycoflora or fungal species diversity as compared to field region without fungal diversity. The fungi conservation is the basic aspect of this investigation. Mycoflora provides fertility to soil and improves its quality increasing the yield of wheat (Triticum aestivum) crop. KEYWORDS: mycoflora, soil, diversity, Kota, wheat, crop, spore, species, fungi, growth, yield How to cite this paper: Amita Mali | Dr. Neerja Shrivastava "Soil Mycofloral Diversity of Wheat Crop Field of Kota Region" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456- 6470, Volume-6 | Issue-4, June 2022, pp.1049-1054, URL: www.ijtsrd.com/papers/ijtsrd50266.pdf Copyright © 2022 by author(s) and International Journal of Trend in Scientific Research and Development Journal. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0) (http://creativecommons.org/licenses/by/4.0) INTRODUCTION Research was conducted on isolation and identification of soil Mycoflora of wheat fields in Kota region. The soil samples used for the study were collected from different field sites located in Kota where only wheat (Triticum aestivum)crop was grown. IJTSRD50266
  • 2. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1050 The soil sample inoculation was carried was carried out using potato dextrose agar media. A total number of five fungi were isolated Aspergillus niger, Aspergillus fumigatus, Aspergillus oryzae, Rhizopus oryzae and Rhizopus stolonifer. Aspergillus fumigatus and Aspergillus niger had the highest percentage frequency of occurrences of 33.33%, Rhizopus oryzae and Rhizopus stolonifer had the lowest percentage frequency of occurrence of 6.67% respectively. Conclusively the amount of work already accomplished in the investigating the micro flora of the soil it must be admitted that no one has yet been able to give a clear picture of all life in the soil and of all interaction of different groups of living things. Developing novel methodologies to study soil ecology under the natural condition in wheat fields is needed and will required collaboration between plant biologists, ecologists, and soil scientist to develop the a system where biochemical and molecular biology studies could be performed on site. It has been estimated that one gram of surface soil contain 50,000 to a million fungi, the loss of organic material from root provides the energy for the development of active fungal population in the Rhizosphere around the root of wheat crop. Wheat (Triticum aestivum) crop field in Kota Methodology The soil samples from wheat fields of Kota region were introduced into a test tube containing 9ml of sterile distilled water. Suspension prepared was labeled and vigorously shaken. Serial dilution was prepared by pippeting 1ml from stock suspension and inoculates in to 9ml of sterile distilled water in a labeled test tubes and shaking vigorously to obtain 10-9 dilution 1ml was further pipette from the first dilution and introduced in to another labeled test tube to obtain 10-1 dilution. They were repeated done to obtain up to 10-9 dilution 0.1ml each from 10-4 and 10-5 dilutions were inoculated in PDA plates and swabbed to ensure even distribution of inoculums in the medium. Also 0.5ml each was taken directly from the stock suspension and inoculated into PDA agar plate and swabbed as above. All plate were labeled and incubated at room temperatures for 5 to 6 days. After incubation the fungal colonies will be developed Fully developed colonies with different cultural characteristic were subculture using sterile inoculating needles in to freshly prepared PDA plates. Each colony was considered to have originated from one fungal propagules Sub cultured plates were incubated at room temperatures for 3–5 days to obtain pure cultures of the various fungal types.
  • 3. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1051 Fungal colonies in petriplates Placing a drop of fungal stain using mounting needle on the clean slide and carefully teased (spread). A drop of lacto phenol cotton blue was added on fungal stain and cover slip was placed on top. Then, examine under microscope using x40 objectives The fungi were identified by way of microscopic method Pure fungal cultures in test tubes Discussion and Results The fungal species isolated and identified are described in table below:- Table-1: Morphological and Cultural Characteristic of the Isolated Fungi Fungal Isolate Colony Texture Aspergillus niger Colonies consisting of a compact white yellowish with dense of dark brown to black. Aspergillus fumigatus Colonies consisting of a dense felt of dark green conidiophores intermixed with aerial hyphae Rhizopus stolonifer Colonies whitish becoming-greenish due to the brownish sporangiosphores and brown black sporangia. Rhizopus oryzae Colony whitish becoming-gray with age, about 10mm high. Stolons smooth or slightly rough almost colorless almost too yellowish brown. Aspergillus oryzae Colonies consisting of a felt of long conidiophores often intermixed with aerial mycelium. Pale greenish yellow, later becoming light to dull brown.
  • 4. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1052 Aspergillus niger Hyphae are septate and hyaline. Conidial heads are radiate initially, splitting into columns at maturity. The species is biseriate (vesicles produces sterile cells known as metulae that support the conidiogenous phialides). Conidiophores are long (400-3000 µm), smooth, and hyaline, becoming darker at the apex and terminating in a globose vesicle (30-75 µm in diameter). Metulae and phialides cover the entire vesicle. Conidia are brown to black, very rough, globose, and measure 4-5 µm in diameter Aspergillus fumigatus Aspergillus colonies are downy to powdery in texture. The surface color may vary depending on the species. The reverse is uncolored to pale yellow in most of the isolates. Aspergillus fumigatus is a thermotolerant fungus and grows well at temperatures over 40°C. This property is unique to Aspergillus fumigatus among the Aspergillus species. Aspergillus fumigatus can grow at a temperature range of 20 to 50 °C. Aspergillus oryzae Other microscopic structures include sclerotia, cleistothecia, aleuriconidia, and Hulle cells. These structures are of key importance in identification of some Aspergillus species. Cleistothecium is a round, closed structure enclosing the asci which carry the ascospores. The asci are spread to the surrounding when the cleistothecium bursts. Cleistothecium is produced during the sexual reproduction stage of some Aspergillus species. Aleuriconidium is a type of conidium produced by lysis of the cell that supports it. The base is usually truncate and carries remnants of the lysed supporting cell. These remnants form annular frills at its base. Hulle cell is a large sterile cell bearing a small lumen. Similar to cleistothecium, it is associated with the sexual stage of some Aspergillus species. Rhizopus oryzae Some morphological features, such as the length of rhizoids and sporangiophores, the diameter of sporangia, the shape of columellae, and the size, shape and surface texture of sporangiospores aid in differentiation of Rhizopus species from each other. Rhizopus stolonifer Nonseptate or sparsely septate broad hyphae (6-15 µm in diameter), sporangiophores, rhizoids (root-like hyphae), sporangia, and sporangiospores are visualized. Sporangiophores are brown in color and usually unbranched. They can be solitary or form clusters. Rhizoids are located at the point where the stolons and sporangiophores meet. Sporangia (40-350 µm in diameter) are located at the tip of the sporangiophores. They are round with flattened bases.
  • 5. International Journal of Trend in Scientific Research and Development @ www.ijtsrd.com eISSN: 2456-6470 @ IJTSRD | Unique Paper ID – IJTSRD50266 | Volume – 6 | Issue – 4 | May-June 2022 Page 1053 Apophysis is absent or rarely apparent and columellae are hemispherical. Sporangiospores (4-11 µm in diameter) are unicellular, round to ovoid in shape, hyaline to brown in color, and smooth or striated in texture Table 2 Frequency of Occurrences of the Isolated Fungi Fungal Isolate Number of Occurrences Aspergillus niger 5 Aspergillus fumigatus 5 Aspergillus oryzae 3 Rhizopus oryzae 1 Rhizopus stolonifer 1 TOTAL 15 Table 3 Percentage Frequency of Occurrences of the Isolated Fungi Fungal Isolate Percentage of occurrences (%) Aspergillus fumigatus 33.33% Aspergillus niger 33.33% Aspergillus oryzae 20.00% Rhizopus oryzae 6.67% Rhizopus stolonifer 6.67% Total Percentage 100% The result obtained in this research indicated that Aspergillus fumigatus and Aspergillus niger has the higher percentages and high number of occurrences. The highest number of fungal species isolated from the rhizosphere of wheat crop is not surprising, due to the production of substrate by growing root in the form of root exudates containing amino, sugar, organic acid, nucleotide and other substrate. Thus, this lead to their proximity to the roots and subsequent proliferation and multiplication of the mycoflora in the soil region. Also high rate of microbial decomposition of both organic and sloughed off tissue (wheat) is yet another factor of their abundance compared with the field soil where such activities are minimal. The fungal flora acts as universal agent of decay, from which new life continually arises and is nourished, the fungi makes a unique contribution to the maintenance of soil fertility. Apart from the saprophytic fungi which act as scavengers, a number of important plant parasitic fungi live in the soil and attack economically, important plants through their roots or at the ground level. The obligate saprophytic fungi isolated were genera of Aspergillus which affect the wheat crop only when they are kept during storage period. Also they add to the total biomass of fungi in the soil and advantageous in the decomposition of organic matter which enhances soil fertility. It was also noted that, the intense microbial activity in the soil (i.e. from both harmful and beneficial microbes) can be especially important because it may result in suppression or even elimination of photogenic microorganism. Rhizopus oryzae affects the wheat plants rottening it. Conclusion The result of this study indicated that five fungal isolate where isolated which include the Aspergillus niger, Aspergillus fumigatus, Aspergillus oryzae, Rhizopus oryzae and Rhizopus stolonifer, which indicate that out of these fungal isolate Aspergillus niger and Aspergillus fumigatus has the higher percentage frequency of occurrence of 33.33%, followed by Aspergillus oryzae which has 20.00%, while the Rhizopus oryzae and Rhizopus stolonifer has the least percentage frequency of occurrence of 6.67% respectively. Thus it can be understood that fungi are also necessary evils. They can be bound in rhizosphere of crop and create fertility in field and can also simultaneously be parasitic to wheat plants. Hence useful fungi should be added to the field and conservation should be done whereas harmful parasitic fungi destroying crops should be treated using antifungals of specific types. Mycoflora in fields of wheat or any crop thus in general decay and make soil fertile making growth and development of plants more effective and in high frequency of fungal species the rate of growth of crops also increases. Hence positive utilities of mycoflora in soil makes farmers to add good saprophytic decaying species in fields in Kota region References [1] Gow, Neil A. R.; Latge, Jean-Paul; Munro, Carol A.; Heitman, Joseph (2017). "The fungal cell wall: Structure, biosynthesis, and function". Microbiology Spectrum. 5 (3). doi:10.1128/microbiolspec.FUNK-0035-2016. hdl:2164/8941. PMID 28513415. [2] Mihail JD, Bruhn JN (November 2005). "Foraging behaviour of Armillaria rhizomorph systems". Mycological Research. 109 (Pt 11): 1195–207. doi:10.1017/S0953756205003606. PMID 16279413. [3] Keller NP, Turner G, Bennett JW (December 2005). "Fungal secondary metabolism - from biochemistry to genomics". Nature Reviews. Microbiology. 3 (12): 937–47. doi:10.1038/nrmicro1286. PMID 16322742. S2CID 23537608. [4] Wu S, Schalk M, Clark A, Miles RB, Coates R, Chappell J (November 2006). "Redirection of cytosolic or plastidic isoprenoid precursors elevates terpene production in plants". Nature
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