INVESTIGATING THE In vitro REGENERATION VIGOUR OF Brassica juncea VAR. VARUNA AND ITS WHITE RUST TOLERANT MUTANT GENOTYPES: CONJUGATING THE CONVENTIONAL AND BIOTECHNOLOGICAL APPROACHES

Main Article Content

ANUPRIYA CHATTERJEE
https://orcid.org/0000-0002-0152-0978
SHRADHA NIRWAN
https://orcid.org/0000-0002-5312-5216
PANKAJ SHARMA
https://orcid.org/0000-0001-7405-1706
ABHA AGNIHOTRI
https://orcid.org/0000-0002-2423-3058
NEERAJ SHRIVASTAVA
https://orcid.org/0000-0003-0092-0184

Abstract

Indian mustard (Brassica juncea (L.) Czernj. Cosson) is an economically significant oilseed crop across the globe with well-known nutritive values. It ranks third among the oilseed crops in the world and second in India. White rust, caused by Albugo candida, is a devastating disease of Brassicaceae family impeding the yield of Indian mustard by up to sixty percent worldwide. One way to overcome this problem is to develop transgenic resistant varieties. Different genotypes in Brassica are known to require different protocols for efficient regeneration protocol which becomes a prerequisite for taking up any genetic improvement plan through transgenic approaches. In this study, we evaluated the regeneration potential of B. juncea check white rust susceptible variety Varuna and its two white rust tolerant mutant genotypes (C66, C69). Of the various hormonal combinations tested, all the genotypes responded best in the presence of MS+ 0.05mg/l dinitrophenylhydrazine + 1.0 mg/l 6-benzylaminopurine for shoot initiation while further development of shoots were best stimulated by 1.0 mg/l 6-benzylaminopurine and rooting response was best on half strength MS+ 2.0 mg/l Indole-3-butyric acid. The regenerated plants were successfully grown to maturity and selfed seeds were harvested. Henceforth, the efficient regeneration protocol optimized here will help us to further genetically transform the B. juncea var. Varuna and its genotypes with white rust resistance genes deploying durable resistance against A. candida. We believe, this study will widen the scopes of biotechnology for sustaining the Indian mustard yield against white rust.

Keywords:
Brassica juncea, Albugo candida, In-vitro regeneration, white rust resistance, agrobacterium-mediated transformation.

Article Details

How to Cite
CHATTERJEE, A., NIRWAN, S., SHARMA, P., AGNIHOTRI, A., & SHRIVASTAVA, N. (2021). INVESTIGATING THE In vitro REGENERATION VIGOUR OF Brassica juncea VAR. VARUNA AND ITS WHITE RUST TOLERANT MUTANT GENOTYPES: CONJUGATING THE CONVENTIONAL AND BIOTECHNOLOGICAL APPROACHES. PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY, 22(9-10), 122-135. Retrieved from https://ikprress.org/index.php/PCBMB/article/view/5964
Section
Original Research Article

References

Mitra M, Gantait S. Tissue culture-mediated biotechnological advancements in genus Brassica. In. Brassica Improvement. Springer, Cham. 2020;85-107.

Bhanu AN, Srivastava K, Singh RK. Advances in agronomic management in Indian mustard for Eastern Uttar Pradesh. Acta Scientific Agriculture. 2019;3(8): 70-79.

Shekhawat K, Rathore SS, Premi OP, Kandpal BK, Chauhan JS. Advances in agronomic management of Indian mustard (Brassica juncea (L.) Czernj. Cosson): An overview. International J of Agronomy. 2012;181(1):14.

Kumar A, Premi OP, Thomas L. Rapeseed-mustard cultivation in India-an overview. National Research Centre on Canolaseed-Mustard: Bharatpur, India; 2008.

FAO perspective. World agriculture: Towards 2015/2030; 2015.
Available:http://www.fao.org/3/y4252E/y4252e00.htm#TopOfPage
Accessed 8 August 2020

Farooq N, Nawaz MA, Mukhtar Z, Ali I, Hundleby P, Ahmad N. Investigating the In vitro Regeneration Potential of Commercial Cultivars of Brassica. Plants. 2019;8(12): 558.

Chatterjee A, Nirwan S, Bandyopadhyay P, Agnihotri A, Sharma P, Abdin MZ, Shrivastava N. Genomic and molecular perspectives of host-pathogen interaction and resistance strategies against white rust in oilseed mustard. Current genomics. 2020;21(3):179-193.

Soto N, Hernández Y, Delgado C, Rosabal Y, Ortiz R, Valencia L, Borrás-Hidalgo O, Pujol M, Enríquez GA. Field resistance to Phakopsora pachyrhizi and Colletotrichum truncatum of transgenic soybean expressing the NmDef02 plant defensin gene. Frontiers in Plant Science. 2020;11:562.

Swamy BM, Samia M, Boncodin R, Marundan S, Rebong DB, Ordonio RL, Miranda RT, Rebong AT, Alibuyog AY, Adeva CC, Reinke R. Compositional analysis of genetically engineered GR2E “Golden Rice” in comparison to that of conventional rice. J of Agricultural and Food Chemistry. 2019;67(28):7986- 94.

Bhayana L, Paritosh K, Arora H, Yadava SK, Singh P, Nandan D, Mukhopadhyay A, Gupta V, Pradhan AK, Pental D. A mapped locus on LG A6 of Brassica juncea line Tumida conferring resistance to white rust contains a CNL type R gene. Frontiers in Plant Science. 2020;10:1690.

Zhang J, Zhang P, Dodds P, Lagudah E. How target-sequence enrichment and sequencing (TEnSeq) pipelines have catalysed resistance gene cloning in the wheat-rust pathosystem. Frontiers in Plant Science. 2020;11:678.

Hwang HH, Yu M, Lai EM. Agrobacterium-mediated plant transforma-tion: Biology and applications. The Arabidopsis Book. 2017;15:0186.

Halpin C. Gene stacking in transgenic plants–the challenge for 21st century plant biotechnology. Plant Biotechnology J. 2005;3(2):141-55.

Ahmad N, Mukhtar Z. Genetic manipulations in crops: Challenges and opportunities. Genomics. 2017;109(5-6): 494-505.

Baltes NJ, Gil-Humanes J, Voytas DF. Genome engineering and agriculture: Opportunities and challenges. In Progress in molecular biology and translational science. Academic Press. 2017;149:1-26.

Paul MJ, Nuccio ML, Basu SS. Are GM crops for yield and resilience possible?. Trends in Plant Science. 2018; 23(1):10-16.

Tyczewska A, Woźniak E, Gracz J, Kuczyński J, Twardowski T. Towards food security: Current state and future prospects of agro-biotechnology. Trends in Biotechnology. 2018;36(12):1219-1229.

Panjabi-Massand P, Yadava SK, Sharma P, Kaur A, Kumar A, Arumugam N, Sodhi YS, Mukhopadhyay A, Gupta V, Pradhan AK, Pental D. Molecular mapping reveals two independent loci conferring resistance to Albugo candida in the east European germplasm of oilseed mustard Brassica juncea. Theoretical and Applied Genetics. 2010;121(1):137-45.

Prem D, Gupta K, Sarkar G, Agnihotri A. Activated charcoal induced high frequency microspore embryogenesis and efficient doubled haploid production in Brassica juncea. Plant Cell, Tissue and Organ Culture. 2008;93(3):269-282.

Cevik V, Boutrot F, Apel W, Robert-Seilaniantz A, Furzer OJ, Redkar A, Castel B, Kover PX, Prince DC, Holub EB, Jones JD. Transgressive segregation reveals mechanisms of Arabidopsis immunity to Brassica-infecting races of white rust (Albugo candida). Proceedings of the National Academy of Sciences. 2019; 116(7):2767-2773.

Bhalla PL, Singh MB. Agrobacterium-mediated transformation of Brassica napus and Brassica oleracea. Nature Protocols. 2008;3(2):181.

Née Sparrow PAH, Irwin JA. Brassica oleracea and B. napus. In Agrobacterium Protocols. Springer, New York, NY. 2015; 287-297.

Guoliang LI, Lixin YUE, Fei LI, Zhang S, Zhang H, Wei QIAN, Zhiyuan FANG, Jian WU, Xiaowu WANG, Zhang S, Rifei SUN. Research progress on Agrobacterium tumefaciens-based transgenic technology in Brassica rapa. Horticultural Plant J. 2018; 4(3):126-132.

Augustine R, Bisht NC. Targeted silencing of genes in polyploids: Lessons learned from Brassica juncea-glucosinolate system. Plant Cell Reports. 2019;38(1): 51-57.

Sparrow PAC, Dale PJ, Irwin JA. The use of phenotypic markers to identify Brassica oleracea genotypes for routine high-throughput Agrobacterium-mediated transformation. Plant Cell Reports. 2004; 23(1-2):64-70.

Sparrow PAC, Snape JW, Dale PJ, Irwin JA.The rapid identification of B. napus genotypes, for high-throughput transformation, using phenotypic tissue culture markers. Acta Horticult. 2006; 706:239-246.

Sparrow PAC, Townsend TM, Arthur AE, Dale PJ, Irwin JA. Genetic analysis of Agrobacterium tumefaciens susceptibility in Brassica oleracea. Theoretical and Applied Genetics. 2004;108(4):644- 650.

Sparrow PAC, Townsend TM, Morgan CL, Dale PJ, Arthur AE, Irwin JA. Genetic analysis of in vitro shoot regeneration from cotyledonary petioles of Brassica oleracea. Theoretical and Applied Genetics. 2004;108(7):1249-1255.

Nanjundan J, Manjunatha C, Radhamani J, Thakur AK, Yadav R, Kumar A, Meena ML, Tyagi RK, Yadava DK, Singh D. Identification of new source of resistance to powdery mildew of Indian Mustard and Studying Its Inheritance. The Plant Pathology J. 2020;36(2):111.

Pental D, Pradhan AK, Sodhi YS, et al. Variation amongst Brassica juncea cultivars for regeneration from hypocotyl explants and optimization of conditions for Agrobacterium-mediated genetic transformation. Plant Cell Reports. 1993;12:462–467.

Pua EC, Chi GL. De novo shoot morphogenesis and plant growth of mustard (Brassica juncea) In vitro in relation to ethylene. Physiol. Plant. 1993; 88:467–474.

Burnett L, Arnoldo M, Yarrow S, Huang B. Enhancement of shoot regeneration from cotyledon explants of Brassica rapa ssp. oleifera through pretreatment with auxin and cytokinin and use of ethylene inhibitors. Plant Cell, Tissue and Organ Culture. 1994;37(3):253-256.

Chi GL, Barfield DG, Sim GE, Pua EC. Effect of AgNO3 and aminoethoxyvinylglycine on In vitro shoot and root organogenesis from seedling explants of recalcitrant Brassica genotypes. Plant Cell Reports. 1990;9(4): 195-8.

Zhang FL, Takahata Y, Xu JB. Medium and genotype factors influencing shoot regeneration from cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis). Plant Cell Reports. 1998; 17(10):780-6.

Eapen S, George L. Plant regeneration from peduncle segments of oil seed Brassica species: influence of silver nitrate and silver thiosulfate. Plant cell, tissue and organ culture. 1997;51(3): 229-32.

Akasaka-Kennedy Y, Yoshida H, Takahata Y. Efficient plant regeneration from leaves of rapeseed (Brassica napus L.): the influence of AgNO3 and genotype. Plant Cell Reports. 2005;24(11):649-54.

Gupta K, Prem D, Agnihotri A. Role of biotechnology for incorporating white rust resistance in Brassica species. In. Plant Biotechnology and Molecular Markers. Springer, Dordrecht. 2004;156-168.

Ono Y, Takahata Y, Kaizuma N. Effect of genotype on shoot regeneration from cotyledonary explants of rapeseed (Brassica napus L.). Plant Cell Reports. 1994 ;14(1):13-7.

Agnihotri A, Gupta K, Prem D, Sarkar G, Mehra VS, Zargar SM. Genetic enhancement in rapeseed-mustard for quality and disease resistance through In vitro techniques. Proceedings of 16th Australian research assembly on Brassicas, Ballarat, Australia. 2009;28.

Cassells AC. Robert N. Trigiano and Dennis J. Gray (eds): Plant Tissue Culture Concepts and Laboratory Exercises, Boca Raton, USA: CRC Press. 2000;454. UK£ 53.99. Plant Cell, Tissue and Organ Culture. 2001; 65(2):174-174.
ISBN: 0-8494-2029-1.

Naeem I, Munir I, Durrett TP, Iqbal A, Aulakh KS, Ahmad MA, Khan H, Khan IA, Hussain F, Shuaib M, Shah AA. Feasible regeneration and agro bacterium-mediated transformation of Brassica juncea with Euonymus alatus diacylglycerol acetyltransferase (EaDAcT) gene. Saudi J of Biological Sciences. 2020;27(5):1324-1332.

Yang ZH, Jin H, Plaha P, Bae TW, Jiang GB, Lim YP, Lee HY. An improved plant regeneration protocol using cotyledonary explants from inbred lines of Chinese cabbage (Brassica rapa ssp. pekinensis). J Plant Biotech. 2004;6: 1-5.

Bhuiyan MS, Min SR, Liu JR. High frequency plant regeneration from transverse thin cell layers in Indian mustard (Brassica juncea L.). Journal of Plant Biotechnology. 2009;36(1):81.

Saeed W, Naseem S, Gohar D, Ali Z. Efficient and reproducible somatic embryogenesis and micropropagation in tomato via novel structures- Rhizoid Tubers. PloS One. 2019;14(5): 0215929.

Kashyap S, Tharannum S, Taarini R. Influence of formulated organic plant tissue culture medium in the shoot regeneration study of Brassica juncea (l.)-Indian mustard. J of Plant Biotechnology. 2019;46(2):114-118.