Breeding Strategies of Mustard for Biotic Stress Resilience By Achyuta Basak.pptx

Biotic Stress Factors Affecting Mustard
Pests:
Aphid (Lipaphis erysimi):
• These small, soft-bodied insects feed on the sap of the plants, causing direct damage to the leaves, stems, and flowers. They also transmit various viral diseases that can further impact plant growth and yield.
• Some common symptoms of aphid infestation in mustard plants include stunted growth, wilting, yellowing or curling of leaves, and the presence of sticky honeydew on the leaves. These symptoms can lead to significant yield losses, with severe infestations causing reductions in seed production, oil content, and overall crop quality.
• Aphids can also cause indirect damage to mustard crops by attracting other pests such as ants, which can further damage the plants. In addition, the honeydew secreted by aphids can lead to the growth of sooty mold, which can further reduce the plant's ability to photosynthesize and produce food.
Diamondback moths (Plutella xylostella):
• These small, grayish-brown moths lay their eggs on the leaves of mustard plants, and the larvae feed on the leaves, causing extensive damage.
• Whitish patches due to scrapping of epidermal leaf tissues by young larvae
• The leaves give a withered appearance but in later stages larvae bore holes in the leaves
• It also bores into pods and feeds developing seed Diamondback moths can also transmit various viral diseases, including turnip mosaic virus, which can further impact plant growth and yield.
Mustard Saw Fly (Athalia lugens proxima):
Symptoms of damage:
• Initially the larva nibbles leaves, later it feeds from the margins towards the midrib
• The grubs cause numerous shot holes and even riddled the entire leaves by voracious feeding
• They devour the epidermis of the shoot, resulting in drying up of seedlings and failure to bear seeds in older plants
Identification of the pest:
• Larva: Greenish black with wrinkled body and has eight pairs of pro-legs. On touch the larva falls to ground and feigns death
• Adult: Head and thorax is black in colour, abdomen is orange colour, wings are translucent, smoky with black veins.
Disease:
Alternaria blight (Alternaria brassicae):
• Alternaria blight is a fungal disease that can significantly impact mustard production. This disease is caused by the pathogen Alternaria brassicae, which infects the leaves, stems, and pods of mustard plants.
• Three species of Alternaria, viz., A. brassicae (Berk.) Sacc., A. brassicicola (Schw.) Wilts. and A. raphani Groves and Skolko have been found to affect the rapeseed and mustard crop quite commonly throughout the world.
• Some common symptoms of alternaria blight in mustard plants include small, circular, brown or black spots on the leaves, which can later coalesce and form large patches of necrotic tissue. The infected plant parts may also become wilted, defoliated, or malformed. These symptoms can lead to significant yield losses, with severe infections causing reductions in plant growth, seed production, and overa

1. Breeding Strategies of Mustard for Biotic Stress Resilience Presented by: Achyuta Basak Reg No- A-2020-020-D Department of Genetics & Plant Breeding Name of the Chairman: Dr. Moumita Chakraborty Seminar Leader: Dr. Shubhrajyoti Sen

2. Mustard: What is it? Scientific name : Brassica juncea Family : Brassicaceae Local name : Rai or Raya or Laha or banga sarson Chromosome No. : (2n=4x=36, genome AABB) The “Triangle of U”, showing the genetic relationships between the six species of the genus Brassica. Brassica juncea is the most predominant crop out of Rapeseed-mustard crops in India and accounts for more than 90% of the area. (Source: www.drmr.res.in ) Its cultivation, which was confined to the Northern belt earlier has now spread to non-traditional areas in Eastern, Western and Southern regions of the country. Mustard ( Brassica juncea ) is the predominant winter oilseed crop in India. Average productivity (1511 kg/ha) of mustard in India is much lower than the world average of 1980 kg/ha for rapeseed-mustard crops ( SOPA 2020 ). Brassica juncea is an amphidiploid (2n=4x=36, genome AABB) derived from interspecific cross of Brassica nigra (2n=2x=16, genome BB) and Brassica rapa (2n=2x=20, genome AA).

3. Indian Mustard vs Toria Sl No. Particular Brassica juncea Brassica rapa 1 Plant Height B. Juncea is a relatively tall plant than Toria . Toria is a relatively shorter plant than junea & having bushy apperance .   2 Cultivation B. Juncea is grown in many parts of the world, including India, China, and Canada Toria is more commonly grown in India and other parts of South Asia 3 Leaves B. Juncea leaves are relatively larger than Toria leaves. Toria leaves are is a relatively smaller than junea leaves. 4 Edibility Both plants are used for their seeds and oil, but B. Juncea is not typically consumed as a leafy vegetable because of bitterness Apart from seed and oil, Toria is also commonly used as a leafy vegetable (known as sarson ka saag in Indian cuisine).   5 Seed Pods B. Juncea has less seed per pod (siliqua) than Toria . Toria has relatively more seed per pod than brassica. 6 Maturity Generally, B. Juncea can reach its maturity stage at 100-120 DAS. But generally, Rye is used to take time for maturity is 80-90 DAS. 7 Seed Colour Mainly the black in colour, sometime it may be yellow also Mainly yellow in colour  Origin & Distribution of Brassica rapa (Source: Royal Botanical Garden: Kew https://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:279485-1 Origin & Distribution of Brassica juncea (Source: Royal Botanical Garden: Kew https://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:60442520-2#distributions

4. Area & Production of Mustard Contribution of different oilseed crops area Average (2014-15 to 2018-19) Contribution of different oilseed crops Production Average (2014-15 to 2018-19) Contribution of different oilseed producing states in total area (2014-15 to 2018-19) Contribution of different oilseed producing states in total production (2014-15 to 2018-19) Source: ICAR-Directorate of Rapeseed-Mustard Research http://www.drmr.res.in/about_rmcrop.php

5. Rapeseed-Mustard Production trends in India Area & Production of Mustard Source: ICAR-Directorate of Rapeseed-Mustard Research http://www.drmr.res.in/about_rmcrop.php

6. Biotic Stress Factors Affecting Mustard

7. Biotic Stress Factors Affecting Mustard: Pests Aphid ( Lipaphis erysimi ) : These small, soft-bodied insects feed on the sap of the plants, causing direct damage to the leaves, stems, and flowers. They also transmit various viral diseases that can further impact plant growth and yield. Some common symptoms of aphid infestation in mustard plants include stunted growth, wilting, yellowing or curling of leaves, and the presence of sticky honeydew on the leaves. These symptoms can lead to significant yield losses, with severe infestations causing reductions in seed production, oil content, and overall crop quality. Aphids can also cause indirect damage to mustard crops by attracting other pests such as ants, which can further damage the plants. In addition, the honeydew secreted by aphids can lead to the growth of sooty mold, which can further reduce the plant's ability to photosynthesize and produce food.

8. Diamondback moths ( Plutella xylostella ): These small, grayish-brown moths lay their eggs on the leaves of mustard plants, and the larvae feed on the leaves, causing extensive damage. Whitish patches due to scrapping of epidermal leaf tissues by young larvae The leaves give a withered appearance but in later stages larvae bore holes in the leaves It also bores into pods and feeds developing seed Diamondback moths can also transmit various viral diseases, including turnip mosaic virus, which can further impact plant growth and yield.

9. Mustard Saw Fly ( Athalia lugens proxima ): Symptoms of damage: Initially the larva nibbles leaves, later it feeds from the margins towards the midrib The grubs cause numerous shot holes and even riddled the entire leaves by voracious feeding They devour the epidermis of the shoot, resulting in drying up of seedlings and failure to bear seeds in older plants Identification of the pest: Larva: Greenish black with wrinkled body and has eight pairs of pro-legs. On touch the larva falls to ground and feigns death Adult: Head and thorax is black in colour , abdomen is orange colour , wings are translucent, smoky with black veins

10. Biotic Stress Factors Affecting Mustard: Disease Alternaria blight is a fungal disease that can significantly impact mustard production. This disease is caused by the pathogen Alternaria brassicae , which infects the leaves, stems, and pods of mustard plants. Three species of Alternaria, viz., A. brassicae (Berk.) Sacc ., A. brassicicola ( Schw .) Wilts . and A. raphani Groves and Skolko have been found to affect the rapeseed and mustard crop quite commonly throughout the world. Some common symptoms of alternaria blight in mustard plants include small, circular, brown or black spots on the leaves, which can later coalesce and form large patches of necrotic tissue. The infected plant parts may also become wilted, defoliated, or malformed. These symptoms can lead to significant yield losses, with severe infections causing reductions in plant growth, seed production, and overall crop quality. Alternaria blight can spread rapidly, particularly in warm and humid conditions, and can cause significant damage to entire mustard fields. It can also reduce the shelf life of harvested mustard seeds and oil. Alternaria blight ( Alternaria brassicae ) :

11. This disease is caused by the pathogen Albugo candida , which infects the leaves, stems, and flowers of mustard plants. Some common symptoms of white rust in mustard plants include small, raised, white pustules on the leaves, stems, and flowers, which can later turn brown and necrotic. The infected plant parts may also become distorted, with twisting or curling of leaves and stems. These symptoms can lead to significant yield losses, with severe infections causing reductions in plant growth, seed production, and overall crop quality. White rust can spread rapidly, particularly in warm and humid conditions, and can cause significant damage to entire mustard fields. In addition, white rust can also reduce the shelf life of harvested mustard seeds and oil. White Rust ( Albugo candida )

12. Black Rot ( Xanthomonas campestris pv . c ampestris ): Black rot is a bacterial disease that can significantly impact mustard production. This disease is caused by the pathogen Xanthomonas campestris pv . campestri s, which infects the leaves, stems, and pods of mustard plants. Some common symptoms of black rot in mustard plants include small, dark, V-shaped lesions on the leaves, which can later turn yellow or brown and become necrotic. The infected plant parts may also become wilted, stunted, or malformed. These symptoms can lead to significant yield losses, with severe infections causing reductions in plant growth, seed production, and overall crop quality. Black rot can spread rapidly, particularly in warm and humid conditions, and can cause significant damage to entire mustard fields.

13. Sclerotinia Stem Rot ( Sclerotinia sclerotiorum ): Symptom The stems develop water-soaked spots which later may be covered with a cottony white growth. As the disease progresses, affected portions of the stem develop a bleached appearance, and eventually the tissues shred. Girdling of the stem results in premature ripening and in lodging of plants. Hard black bodies, the sclerotia, are formed inside the stem and occasionally on the stem surface. Basal stalk infections rarely occur. Yield loss of 10 to15% has occurred in Saskatchewan, Manitoba and North Dakota; occasionally losses of 50% have occurred in Manitoba

14. Black Leg ( Leptosphaeria maculans ) It is caused by a hemi-biotrophic fungus Leptosphaeria maculans , belonging to the class Ascomycetes. The disease is prevalent in Australia, Canada, and Europe. Symptoms: The disease appears as white/buff-colored lesions on the leaves and white or grey lesions with a dark border on stems. Under severe infection conditions, the fungus girdles the stem at the crown thereby leading to the lodging of the plant. Blackleg infection on pods results in premature pod shattering and seed infection (Raman et al. 2013 ). It germinate in the presence of free water from 4-28ºC (40-82ºF). Penetration is through stomates. The pathogen also may be seedborne. Seeds may be infected and/or infested by the pathogen.

15. Biotic Stress Factors Affecting Mustard: Weeds Some common symptoms of weed infestation in mustard fields include reduced plant growth and vigor, uneven stand establishment, and reduced seed production. Weeds can also interfere with the harvesting process and reduce the efficiency of post-harvest operations. Weeds can have a significant impact on mustard production, with yield losses ranging from 10% to 80% depending on the severity of weed infestation and the weed species present. Example: Bathua ( Chinnapodium album ), Bermuda grass ( Cynodon dactylon ), Kharthua ( Chinnapodium murale ), Shepherd’s purse ( Capsella bursa-pastoris ), Gehul ka mama ( Phaliris minor ) etc…

16. Biotic Stress Factors Affecting Mustard : Nematodes Root-knot nematodes ( Meloidogyne spp .) Reniform nematodes ( Rotylenchulus spp .) Lance nematodes ( Hoplolaimus spp .) Sting nematodes ( Belonolaimus spp .) Spiral nematodes ( Helicotylenchus spp.) Lesion nematodes ( Pratylenchus spp .) Cyst nematodes ( Heterodera and Globodera spp .) Plant parasitic nematodes are microscopic roundworms that can significantly impact mustard production. These pests feed on the roots of mustard plants, causing damage to the root system and reducing plant growth, yield, and quality. Some common symptoms of nematode infestation in mustard plants include stunted growth, reduced plant vigor, yellowing or chlorosis of leaves, and poor root development. These symptoms can lead to significant yield losses, with severe infestations causing reductions in plant growth, seed production, and overall crop quality. Plant parasitic nematodes can spread rapidly in soil and can cause significant damage to entire mustard fields. In addition, nematode infestations can also increase the susceptibility of mustard plants to other biotic and abiotic stresses, such as drought, salinity, and diseases.

17. T he prevalence and distribution of biotic stress in mustard production The prevalence and distribution of biotic stress can vary depending on the region, cropping system, and season. In India, which is the largest producer and consumer of mustard globally, several biotic stresses can affect mustard production. The most common pests that affect mustard in India include aphids, flea beetles, and diamondback moth. Diseases such as white rust, black rot, and A lternaria blight can also cause significant yield losses in mustard crops. Weeds such as wild oats, wild mustard, and shepherd's purse are also prevalent in Indian mustard fields. In addition to India, mustard production in other regions such as China, Canada, and Australia is also affected by biotic stresses. For instance, flea beetles and diamondback moth are major pests that affect mustard production in Canada, whereas blackleg and white rust are significant diseases that affect mustard crops in Australia. The prevalence and distribution of biotic stress can also vary depending on the season. For example, flea beetles and diamondback moth are more prevalent during the early stages of mustard growth in the spring, whereas aphids and white rust are more common during the later stages of growth in the fall.

18. Objectives of Mustard Breeding for Biotic Stress Resilience Developing plants with improved nutritional quality that can better withstand biotic stress and support human health Improved nutritional quality Developing plants that can resist or tolerate biotic stresses caused by pests, diseases, and other organisms Resistance to biotic stress Developing plants with a diverse genetic pool to increase resilience against a range of biotic stressors Genetic diversity Developing plants that can adapt to changing environments and biotic stressors, such as those caused by climate change Adaptability Developing plants that can maintain high yields in the face of biotic stress. Yield stability

19. Traditional Breeding Approaches for Biotic Stress Resilience: Mass Selection 06 - Pusa Mustard-29 high yield potential and resistance to Alternaria blight 01 - Pusa Bold High yield potential and resistance to aphids, white rust, and Alternaria blight. 04 - Pusa Mustard-30 high yield potential and resistance to Alternaria blight. 03 - Pusa Mustard-21 high yield potential and resistance to white rust. 05 - Pusa Mustard-28 high yield potential and resistance to white rust. 02 -Pusa Basanti high yield potential and resistance to white rust. Mass selection

20. Pedigree Selection & Recurrent Selection

21. Hydridization & backcross breeding

22. Modern Breeding Approaches for Biotic Stress Resilience Zhang et al. (2019 ) used high-throughput genotyping-by-sequencing technology to identify molecular markers associated with resistance to Fusarium wilt disease in mustard. They identified several candidate genes and markers that can be used for marker-assisted breeding to develop Fusarium wilt-resistant mustard varieties. Assou et al ( 2022 ) Remove the major allergen Bra j I, a seed storage protein of the 2S albumin class, in the allotetraploid brown mustard ( Brassica juncea ) using genome editing via CRISPR/Cas9 NRCHB-101 (Narendra Rai Chaudhary Mustard Hybrid 101). It is a hybrid variety developed by scientists at the Indian Council of Agricultural Research (ICAR) using gene stacking for disease resistance. Different types of Omics technology been used for biotic stress resilience of Mustard- Association Mapping, Introgressive breeding, NGS-based BSA, Transcriptomics and Proteomics. Omics Technologies CRISPR-Cas9 Technology . High-throughput sequencing. Gene stacking .

23. Different disease resistance QTLs identified in Brassica juncea Population used Markers used Mapped gene/QTLs References White rust Disease Donskaja × Jubilejnaja F1 derived DH RFLP Acr = Ac2a1, monogenic dominant gene (race 2A) Cheung et al. ( 1998) BEC144 X Varuna (F7 ; F2 ) RIL populations RAPD, CAPS, AFLP AC2 (t) Mukherjee et al. (2001) , Varshney et al. ( 2004) Heera X Varuna (F1 Derived DH) AFLPs, SSRs AcB1-A4.1 Panjabi- Massand et al. ( 2010 ) Donskaja IV X TM-4 AFLPs, SSRs AcB1-A5.1 Panjabi- Massand et al. ( 2010 ) Tumida X Varuna F1 derived DH SSR, SNP BjuWRR2 Bhayana et al. ( 2020 )

24. Different disease resistance QTLs identified in Brassica juncea Population used Markers used Mapped gene/QTLs References Scle r otinia stem r ot B. juncea —B. fruticulose Introgression lines SNP and SSR A01, A03, A04, A05, A08, A09, and B05 Rana et al . ( 2017 ) B. juncea —B. fruticulose Introgression lines GBS and GWAS A01, A03, A04, A05, A08, A09 and B05 ad 20 candidate genes Atri et al . ( 2019 ) B. juncea —E. cardamonides Introgression lines GBS and GWAS A03 and B03 Rana et al. ( 2019 ) Bla c kl e g Recombinant Lines ( B. napus and B. juncea ) RAPD, RFLP B8 (Rlm 6 ) Ch e vre et al. ( 1997 ) A C V ulcan/UM3132 (F2 ) RFL P , SSR LMJR1 Christianson et al. ( 2006 ) A C V ulcan/UM3132 (F2 ) RFL P , SSR LMJR2 Christianson et al. ( 2006 ) Different disease resistance QTLs identified in Brassica juncea

25. 02 Introgressive breeding Incorporate useful genes into cultivated popular varieties from wild species, has been exploited for transferring disease resistance loci from exotic and wild relatives Atri et al . (2019) mapped resistance responses against stem rot on seven B. juncea LGs: A01, A03, A04, A05, A08, A09, and B05 through GWAS in the set of introgression lines of B. juncea —B. fruticulosa 03 NGS-Based Bulked Segregant Analysis In this technique, DNA or RNA from contrasting segregated phenotypes are bulked to form pools. These pools were genotyped followed by detection of QTLs through SNPs calling (Liu et al . 2012 ; Takagi et al . 2013 ) NGS based BSA has been used for fine mapping and cloning of the blackleg resistance gene, Rlm1 , in B. napus . Rlm 1 shares homology with STN7 (B. rapa , B. oleracea , and Arabidopsis) encoding a serine/threonine-protein kinase which is involved in triggering the systemic immune response via the production of reactive oxygen species. It encodes a serine/threonine kinase protein (Fu et al . 2019). 01 Association Mapping Mapping of quantitative trait loci that takes advantage of linkage disequilibrium to link phenotypes to genotypes. Raman et al. (2016) mapped Rlm12 on ChrA01 using a B. napus diversity panel comprising of 179 lines. Additionally, they identified the previously mapped Rlm4 on A07, and several new SNPs showing a strong association with the blackleg resistance trait (Raman et al. 2020). Application of the Omics Technologies in Brassica

26. E xamples of mustard varieties developed using different Genetic engineering methods & Marker Assisted selection for biotic stress resistance Braj mustard ( Brassica juncea ) - developed using genetic engineering to express a modified cry1Ac gene from Bacillus thuringiensis ( Bt ) for resistance against the diamondback moth ( Plutella xylostella ) Coral mustard ( Brassica juncea ) - developed using genetic engineering to express the Bt cry1Ac gene for resistance against the diamondback moth Parinay mustard ( Brassica juncea ) - developed using marker-assisted selection to introgress the Bt cry1Ab gene from Brassica napus for resistance against the diamondback moth Surya mustard ( Brassica juncea ) - developed using genetic engineering to express the Bt cry1Ab gene for resistance against the diamondback moth ProAgro mustard ( Brassica juncea ) - developed using genetic engineering to express the Bt cry1Ac and cry1Ba genes for resistance against the diamondback moth and cabbage looper ( Trichoplusia ni ) Pusa Mustard-30 ( Brassica juncea ) - developed using marker-assisted selection to introgress the Sclerotinia resistance gene ( Rlm2 ) from Brassica rapa for resistance against Sclerotinia stem rot RH-30 ( Brassica juncea ) - developed using marker-assisted selection to introgress the clubroot resistance gene ( CRb ) from Brassica rapa for resistance against clubroot Pusa Mustard Hybrid-26 ( Brassica juncea ) - developed using marker-assisted selection to introgress the Sclerotinia resistance gene ( Rlm2 ) from Brassica rapa for resistance against Sclerotinia stem rot NUDHHS-1 ( Brassica napus ) - developed using genetic engineering to express the Bt cry1Ac gene for resistance against the diamondback moth RWG1 ( Brassica juncea ) - developed using marker-assisted selection to introgress the clubroot resistance gene ( CRa ) from Brassica rap a for resistance against clubroot.

27. Transgenic Approaches to Improve Brassica Juncea Biotic Stress Tolerance Cat e gory Genes used Source T ransgenic plants Resistance a g ainst p h ytopathogens/pests References Signaling path w ays B j NPR1 B. juncea B. juncea A. b r assica e , Erysiphe cruciferarum Ali et al. ( 2017 ) Germin and germin like proteins HvOxO H. vulga r e B. juncea Sclerotinia sclerotiorum V erma and Kaur ( 2021 ) PR proteins Chitinase, glucanases Solanum lycope r sicum , Barl e y B. juncea A. b r assicae Mondal et al. ( 2007 ), Chhikara et al . ( 2012 ) R genes WRR4 A r abidopsis thaliana B. juncea and B. napus Al b ugo candida Borhan et al . ( 2010 ) WRR1 B r assica juncea B. juncea A. candida Arora et al. ( 2019 ) Lectins H e v ein Rubber t r ee B. juncea A. b r assicae Kanrar et al. ( 2002 ) Agglutinin Colocasia esculenta B. juncea Lipaphis erysimi Das et al. ( 2018 ) Agglutinin A CA Allium cepa B. juncea L. erysimi Hossain et al. ( 2006 )

28. Case Study

29. Rani et al. ( 2017 ) suggested a multi-toxin engineering approach to incorporate aphid resistance in mustard plants. They transformed  B. juncea  cv. Varuna with a fusion gene construct harbor lentil  lectin  gene and chickpea  protease inhibitor  gene under a phloem-specific promoter  rolC  for incorporation of aphid resistance. Since mustard aphid sucks sap from the phloem part of the plant, so phloem-specific expression of the transgene was attempted. In aphid bioassay, the larval survival was inhibited upto an extent of 40% and lesser leaf damage was recorded in transgenic plants than the control plants. 1

30. During the proteomic investigation between these two B. juncea cultivars and R. indica , 9% proteins were identified as stress/defence related proteins. Aphid resistance in mustard cultivars is connected to the level of expression of such proteins. Five isoforms of pentatricopeptide repeat containing protein (PTPR) (Protein no. 23, 26, 116, 130 and 179) were detected during mustard-aphid compatible and incompatible interaction. F26A19.13, a Pentatricopeptide repeat-containing protein, has been found as one of the most essential defence proteins in the plant-insect defence cross-talk because it directly reduces the action of aphid salivary amylase. 2

31. Kamble et al . (2013) developed transgenic B. juncea expressing a synthetic CRY1AC gene using Agrobacterium-mediated gene transfer system to incorporate resistance against Plutella xylostella . The transgene integration was confirmed by Southern hybridization and expression was confirmed by RT- PCR and ELISA of CRY1AC protein. Transgenic mustard plants exhibited significant level of resistance to the 2nd instar larvae of diamondback moth. 3

32. Tasleem et al. ( 2017 ) transformed  B. juncea  with  MPK3  gene and analyzed its role in imparting tolerance against  A. brassicae . The estimation of activity of antioxidative enzyme was determined in transgenic plants. It was found that both ascorbate peroxidase (APX) and guaiacol peroxidase activity as well as proline content were higher in transgenic plants, which led to scavenging of ROS in transgenic plants produced as a result of  Alternaria  infection, which is responsible for inducing resistance trait in transgenic mustard plants. 4

33. They identified and isolated a CC-NB-LRR protein-coding  BjuWRR1  gene from Donskaja -IV, an exotic line of Indian mustard and transferred it into Indian mustard by  Agrobacterium -mediated gene transfer technique to validate its role in providing resistance against white rust pathogen,  Albugo candida . Transgenic mustard plants exhibited resistance against six isolates of white rust fungus collected from different Indian mustard growing zones of India. 5

34. The study is based on crop preference of Plutella xylostella by testing 5 different plants i.e. cabbage, cauliflower, broccoli, Brassica pekinensis (Lour.) and Indian mustard, Brassica juncea (L.) Czern & it is found that females preferred to oviposit on Indian mustard. It is also found that low larval survival on Indian mustard in the laboratory, and low infestations in the field, as compared to the high oviposition preference in the laboratory of P. xylostella moths on Indian mustard, can be recommended as a trap crop in South Africa. 6

35. Deployment and Adoption of Mustard Genotypes with Biotic Stress Resilience Deployment involves the distribution and promotion of the new genotypes to farmers. Adoption, on the other hand, involves farmers actually using the new genotypes in their fields. Farmers may be hesitant to adopt new varieties In the case of mustard genotypes with biotic stress resilience, successful deployment and adoption could lead to increased crop yields and reduced dependence on chemical inputs, resulting in improved sustainability and profitability for farmers. It could also contribute to the broader goal of global food security by increasing the availability and accessibility of a key crop.

36. Challenges of Mustard Breeding for Biotic Stress Resilience Genetic complexity Limited genetic resources Regulatory frameworks High costs of modern breeding technologies Climate change Resistance development

37. Conclusion & Future Prospects of Mustard Breeding for Biotic Stress Resilience Since major yield losses occur due to biotic stresses imposed on the crop, therefore, disease management and improving resistance are the top priorities in the breeding programs. However, achieving the goals for stress resilience will require overcoming several challenges, & to mitigate this challenges sustained investment in research and development, collaboration among stakeholders across the public and private sectors, and effective communication and engagement with farmers, policymakers, and consumers are essential. In the future, the continued advancement of breeding technologies, such as genome editing and precision breeding, may enable mustard breeders to identify and transfer desirable biotic stress-resistance traits more efficiently and effectively. Moreover, the development and deployment of mustard genotypes with biotic stress resilience may enable farmers to reduce their reliance on chemical pesticides and herbicides, contributing to sustainable agriculture practices and protecting the environment.

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