Exploitation of AVRDC s Chili Pepper (Capsicum spp.) Germplasm in India 1

臺灣園藝 (J. Taiwan Soc. Hort. Sci.) 61(1):1-9, 2015 Exploitation of AVRDC s Chili Pepper (Capsicum spp.) Germplasm in India 1 亞蔬中心辣椒種原在印度的開發與利用 by M.K. Reddy 1, A. Srivastava 2, S.W. Lin 3, R. Kumar 4, H.C. Shieh 3, A.W. Ebert 3, N. Chawda 5, and S. Kumar *3 Additional index words: hot pepper, F1 hybrid, seed sector 附加關鍵字 : 辣椒 雜交一代 種子公司 Abstract: Chili pepper (Capsicum spp.) is an important commercial crop for smallholders in India, where C. annuum L. is the most widely cultivated species for its pungent (chili pepper) and non-pungent (sweet pepper) fruits. During 2012-13, India exported chili pepper and derived products worth about 410 million US $. Over the past 13 years (2001-2013), AVRDC The World Vegetable Center has supplied more than 33000 pepper seed samples of its improved lines (81%) and germplasm accessions (19%) to both public and private institutions in more than 120 countries. India ranked first in terms of acquisition of pepper seed samples (5187) from AVRDC and accounted for 15.6% of the total samples distributed worldwide. The successful uses of supplied germplasm accessions and improved breeding lines in India is described, especially the release and commercialization of open pollinated (OP) and hybrid cultivars. Highly conservative estimates suggest that hybrids involving AVRDC germplasm and improved lines were cultivated on more than 30,000 ha during 2012-13 in different regions of India. 1. Introduction Chili pepper (Capsicum spp.) is an important commercial vegetable and spice crop for smallholder farmers in Asia, Africa and South America. Among the five cultivated species of the genus Capsicum, C. annuum is the most widely cultivated in India for its pungent (chili pepper syn. hot pepper) and non-pungent (sweet pepper syn. capsicum, bell pepper) fruits. The cultivation of C. frutescens L., C. chinense Jacg., and C. baccatum L. is limited and usually restricted to homestead gardening in different regions. According to an 1. Indian Institute of Horticultural Research, Bengaluru 560089, India. 2. Indian Agricultural Research Institute, New Delhi 110012, India. 3. AVRDC The World Vegetable Center, Tainan 74199, Taiwan. 4. Indian Institute of Vegetable Research, Varanasi 221305, India. 5. VNR Seeds, Raipur 492001, India. *Corresponding authors email: sanjeet.kumar@worldveg.org 6. 本文於民國一百零三年十一月二十四日收到 民國一百零四年三月二十七日接受刊登 Received for publication: 24 Nov., 2014. Accepted for publication: 27 Mar., 2015. -1

2- 臺灣園藝第六十一卷第一期 estimate for 2012, chili pepper (dry-red and fresh-green fruits) were cultivated on 801,500 ha in India with a total production of 1.3 million t of dry fruits and 6800 t of fresh fruits. Average yield was 1.6 t ha -1 and 8.5 t ha -1 for dry and green chili, respectively (FAOSTAT, 2012). India is one of the major exporters of dry chili pepper and derived products to different countries (Fig. 1) and during 2012-13 total value of export was about 410 million US$. This value does not include export of fresh chili and chili oleoresin, which constitutes a major share of spice oleoresins exported from India (Reddy et al., 2014). Public sector institutions are mostly involved in strategic research to develop open pollinated varieties (OPVs) and hybrids with limited dissemination efforts of improved seed, while private seed companies are mostly involved in development, promotion and commercialization of hybrid cultivars. Over the past 25 years, AVRDC The World Vegetable Center at its headquarters in Taiwan has been executing pepper breeding project and making its germplasm accessions and improved breeding lines available as international public goods (http://avrdc.org/seed/seeds/). Recently, we published an overview of chili pepper breeding research in India (Reddy et al., 2014). This paper describes successful uses of AVRDC s chili pepper germplasm accessions and improved lines by the Indian researchers and the Indian seed industry in breeding research, cultivar development and commercialization. 2. Indian chili pepper cultivar release process and seed industry In India, the formal chili pepper varietal release procedure is facilitated by All India Coordinated Research Project (AICRP) on vegetable crops (responsible for coordination of multilocation evaluation and recommendation of better performing OPVs or hybrids for release and cultivation) and the Central Variety Release Committee (CVRC) on Horticultural Crops (responsible for official release notification of cultivars). In addition, different state governments also release or notify varieties through State Varietal Release Committees (SVRCs) in respective states. Since its inception in 1971, AICRP has recommended 24 OPVs and 15 hybrid chili cultivars. The CVRC on Horticultural Crops has notified 57 chili pepper cultivars including seven hybrids (all hybrids were notified after 2005) for cultivation in different zones. Under the liberalized Indian seed policy during late 1980s, this formal release process was made non-mandatory for cultivar commercialization. Thereafter, a very sharp growth in Indian vegetable seed sector was achieved and Fig. 1. Indian export share of dry chili and its products (excluding oleoresin and fresh fruits) to different countries. 圖 1. 印度出口乾辣椒以及相關產品 ( 不包括辣椒油與新鮮果實 ) 在不同國家的百分比例

Exploitation of AVRDC s Chili Pepper (Capsicum spp.) Germplasm in India -3 currently there are about 850 small, medium and large-sized seed companies operating in India (Reddy et al., 2014).Out of them, about 50 have R&D facilities many of them with world-class, competitive human and infrastructural resources. Hybrid cultivars account for about 25% of the total area under chili pepper cultivation. About 25% of the area is under improved OPVs and the remaining 50% area is still under local landraces. The current chili pepper hybrid seed market is about 50 t per year, with an estimated turnover worth about 16 million US$. These figures do not include seeds of hot pepper OPVs or sweet pepper OPVs and hybrids (Reddy et al., 2014). 3. Pepper germplasm dissemination by AVRDC The AVRDC Genebank currently has 8235 Capsicum spp. accessions in ex-situ conservation the world s largest collection in the public domain. The Center disseminates seed of its germplasm accessions and improved pepper lines to public and private sector breeders. One of the strategies of AVRDCs pepper breeding program is to disseminate improved pepper lines through the distribution of sets of the International Chili Pepper Nursery (ICPN) and the International Sweet Pepper Nursery (ISPN). As of 2014, 23 ICPN and 12 ISPN nurseries have been released, which included 164 newly developed improved chili (90) and sweet pepper (74) lines. More than 33,000 germplasm materials were distributed from 2001-2013 to more than 120 countries, comprising 81% improved lines and 19% genebank accessions (Kumar et al., 2014). 4. Pepper germplasm acquisition in India from AVRDC Indian public and private sector institutions ranked first in terms of acquisition (between 2001 and 2013) of pepper seed samples (5187) from AVRDC and accounted for 15.6% of the total samples distributed worldwide. The high number of requests and supply of seed samples to India can be explained by the fact that India has the largest number of seed companies and one of the largest public national agriculture research and extension system (NARES) in the world. Nearly 90% of the seed samples received in India were AVRDCs improved breeding and elite lines with traits such as disease resistance and CMS. The largest share of seed samples was acquired by seed companies (62%), NARES (25%) and others (Fig. 2). This high share of Indian seed companies (62%) was almost double the average of seed company recipients in all other countries (33%) (Lin et al., 2013). Fig. 2. AVRDC s peppers (chili and sweet) seed distribution in India: number of samples and percentage by recipient category. 圖 2. 亞蔬的番椒種原 ( 辣椒與甜椒 ) 在印度不同應用者的分布百分比例

4- 臺灣園藝第六十一卷第一期 5. Chili pepper germplasm utilization in India Indian chili breeders from both public and private sectors have used the Center s chili pepper germplasm and improved lines in various ways. Mostly these lines have been: (i) subjected to selection and purification according to local trait preferences and subsequently released as new varieties through national varietal release procedures; (ii) used (possibly after further selection) as parental lines in hybrid development; or (iii) used as sources of traits (such as multiple disease resistance) in crosses to develop new breeding lines. Examples are described in the following sections. 5.1. Use in strategic breeding research AVRDCs identified anthracnose resistant C. baccatum sources (PBC80/VI046804 and PBC81/ VI046805) have been utilized in crosses by breeders to transfer resistance into C. annuum lines. AVRDC s germplasm and improved lines were screened against anthracnose (C. capsici) and resistant sources (CCA4261, C03453 [C. annuum]; C00309 [C. frutescens]) and potential CMS based crosses (CCH-1, CCH-2, CCH-3 and CCH-4) were identified (Garg et al., 2013). Syngenta, India in its international pepper breeding program, has extensively used PBC167 (VI046819) and MC-4 (both obtained from the center) as donor parents in resistance breeding against powdery mildew and bacterial wilt, respectively. The VNR Seeds has developed Phytophthora blight tolerant hybrid using one of the parents from AVRDC (Fig. 3). Recently, potential crosses based on CMS lines introduced from AVRDC have been developed for evaluation (Singh et al., 2014). Likewise, a number of germplasm accessions and improved lines have been used by the Indian breeders (Table 1). 5.2. Use in applied research (cultivars release and commercialization) Since 2005, 12 germplasm accessions and 18 improved pepper lines have been released as 30 OPVs for commercial cultivation in eight countries (Lin et al., 2013). In addition, 34 hybrids developed using AVRDCs materials as parental inbreds were commercialized in six other countries and in India (Fig. 4). The highest number of releases and level of commercialization of cultivars based on AVRDCs germplasm took place in India with a total of 21 hybrids and 2 OPVs (Fig. 4). Most of these hybrids were developed and commercialized by seed companies. One of the populations, KA2 (VI037808) introduced from AVRDC was found to be heterogeneous and breeders at the Indian Institute of Vegetable Research (IIVR) selected highly desirable plants from this population, purified it and increased the seeds (Fig. 5). The purified population was then tested under AICRP and in 2005, based on its better performance over other entries; KA2 was recommended for release and notified in 2006 as Kashi Anmol (Lin et al., 2013). The cultivar became popular among the traders and farmers because of its earliness, short duration (suitable as catch crop under rice wheat cropping system) and desirable fruit size shape and appearance (suitable for traditional packaging in bags made of jute). Other public sector cultivars (based on AVRDC germplasm) recommended for commercial cultivation are Kashi Surkh, Kashi Early, Arka Sweta and Arka Meghna hybrids, and Kashi Sinduri (VRPBC535) OPV for oleoresin extraction (Reddy et al., 2014). Breeders in India used AVRDCs CMS lines for commercial hybrid development, cost-effective seed production, or for converting CMS lines into more preferred genetic backgrounds to develop parents for potential hybrids. For example, Kashi Surkh, and Kashi Tej hybrids

Exploitation of AVRDC s Chili Pepper (Capsicum spp.) Germplasm in India -5 released by IIVR, were developed using AVRDC s CMS lines (Kumar et al., 2007; Kumar et al., 2009); however, these hybrids have not yet been commercialized. Fig. 3. Phytophthora susceptible VNR-435 (left) and tolerant VNR Sunidhi (right) hybrids, a trial plot in Baster (Chhatishgarh), India. 圖 3. 比較在印度 Baster 田間種植雜交一代感疫病辣椒品種 VNR-435 ( 圖左 ) 與耐疫病品種 VNR Sunidhi ( 圖右 ) Table 1. Use of chili pepper germplasm accessions and improved lines from AVRDC in India s breeding research. 表 1. 利用亞蔬的辣椒種原與改良品系應用在印度的育種研究上 Supplied germplasm (alternate name) Utilization status Reference 13 germplasm accessions Used with an array of other germplasm for seed protein diversity analysis PBC535 (VI037556), PBC375 (VI039372), PBC385 (VI039374), PBC66 (VI037518) One pair of CMS (AVPP0711-S), maintainer and a number of other lines Eight different pairs of CMS and their maintainer lines Used to demonstrate that somaclonal variation is genotype dependent Used to study the cytological mechanism of male sterility, and distribution of restoration-of-fertility (Rf) locus associated markers Used to study genetic relationship between two male sterile cytoplasms C. chacoense, KA2 (VI037808) Used to develop interspecific cross and create an alloplasmic male sterile cytoplasm Anu and Peter, 2006 Anu et al., 2004 Kumar et al., 2007 Kumar et al., 2009 Kumar et al., 2009 VI037455, VI037453, VI046889, VI039369, AVPP9807 Identified as resistant sources against Chili veinal mottle virus (ChiMoV) Reddy and Reddy, 2010 A number of germplasm accessions and improved lines Screened against leaf curl disease under field conditions Kumar et al., 2011 PBC535 (VI037556), VI037808 Used to standardize regeneration protocols Kumar et al., 2012 C00304 (VI029136), 337-7545 (AVPP0304), PBC535 (VI037556), CCA-4261(AVPP0711-S), PBC473 (AVPP9623), KA2 (VI037808), 9955-15 (AVPP9905) Used along with highly pungent Indian landraces for marker based genetic diversity analysis Rai et al., 2013 AVPP0102, VI037518, VI037519, VI037548, VI039374, VI037556 Identified as bacterial wilt (caused by Psedomonas solanacearum) resistant sources Reddy et al., 2014 VI037455, AVPP9812 PBC535 (VI037556) Identified as resistant sources against Cucumber mosaic virus (CMV) Used as susceptible parent to study genetics of resistance in Bhut Jolokia to Pepper leaf curl virus Reddy et al., 2014 Rai et al., 2014

6- 臺灣園藝第六十一卷第一期 Fig. 4. India ranked first among 14 countries in releasing and commercializing pepper cultivars 圖 4. 比較番椒發表並商品化的品種數量, 在 14 個國家中以印度為最多者 Fig. 5. Steps involved in selection of KA2 (Kashi Anmol released in 2006) from an AVRDC supplied germplasm (VI037808). 圖 5. 由亞蔬提供的種原 VI037808 KA2 的純化選育過程, 選出的純化品系 ( 商品名 Kashi Anmol) 並於 2006 年在印度發表 The most remarkable commercial successes with AVRDC CMS lines and other improved lines have been achieved by seed companies. For example, VNR Seeds, Chhattisgarh started using AVRDCs improved chili pepper lines in early 2000. Since then, this company has used AVRDC-supplied lines to develop and commercialize hybrids for different market segments in India: VNR-8-7, VNR-Charmee, VNR-G-166 are for drying purposes (Fig. 6), VNR-44-7, VNR-G-Jyothi are for red and green/bhajia (dual) purpose; VNR-38 for pickle processing (Fig. 6); VNR-277 for green fruit export to Middle East. Hybrids based on AVRDC germplasm were also commercialized by other seed companies (Table 2). Highly conservative estimates based on amount of seed sale of hybrids by different seed companies suggest that hybrids involving AVRDC germplasm and improved lines were cultivated on more than 30,000 ha during 2012-13 in different regions of India (Reddy et al., 2014). In addition to profits earned by seed companies, seed distributors, and contract seed producers in a crop year, the adoption of highly profitable hybrid crops benefits an estimated 100,000 small-scale growers, who dominate commercial chili pepper cultivation. It is worthwhile to note that the use of CMS lines reduces the cost of hybrid seed production by at least 50% compared with conventional (manual emasculation) hybrid seed production, which requires skilled labor for manual emasculation and pollination. Skilled labor is more and more difficult to find in India due to migration of laborers to other sectors (Lin et al., 2013).

Exploitation of AVRDC s Chili Pepper (Capsicum spp.) Germplasm in India -7 Fig. 6. Farmers cultivating hybrids VNR-G-166 (suitable for dry fruits) in Kurnul (Andhra Pradesh) and VNR-38 (suitable for pickle processing) in Gondal (Gujarat), India (2012-13). 圖 6. 適合做乾椒品種 VNR-G-166 在 Kurnul (Andhra Pradesh) 與適合做泡椒品種 VNR-38 在 Gondal (Gujarat) 由印度農民在田間栽種情形 (2012-2013) Table 2. Commercialized private sector chili pepper hybrids in India (since 2005) involving AVRDCs germplasm. 表 2. 亞蔬之辣椒種原自 2005 年起已應用在印度種子公司雜交一代辣椒品種上 Name of hybrid (total #) Developed by VNR-8-7, VNR-Charmee, VNR-G-166, VNR-44-7, VNR-Jyothi (VNR-G-273), VNR-38; VNR-277, VNR-332 (Rani), VNR-Vidya (9) Ulka F1, Masaya 315,Yuvraj IN (3) Hybrid (1) NS-222, NS-227, NS-230, NS-638 (4) VNR Seeds, Raipur East-West Seeds, Bengaluru Indus Seeds, Bengaluru Namdhari Seeds, Bengaluru 6. Conclusions Informal tracking of AVRDC germplasm distribution and use suggests that a major multiplying impact of the Center s improved lines and genebank accessions has been achieved in countries like India, where breeder s skills are abundant and improved seed distribution systems are available. Breeding and seed dissemination efforts, along with capacity building of human resources, will remain AVRDC s priority for increasing the reach and impact of the Center s improved pepper lines. Recently, the Center has commissioned a survey to analyze the impact of AVRDC s supplied pepper germplasm in South and Southeast Asian countries. In addition to continuous chili breeding efforts on resistant to major biotic stresses, Indian chili breeding should also focus on post-harvest processing (e.g. oleoresin extraction) and safer chili products (e.g. reduced aflatoxins).

8- 臺灣園藝第六十一卷第一期 Acknowledgements All the authors acknowledge the Indian Council of Agricultural Research (ICAR), New Delhi for providing public research funds to support the Indian chili breeding program. Furthermore, the AVRDC s authors express their thanks to AVRDC s donors for continuous support to the Center. References 1. Anu, A., K.N. Babu and K.V. Peter. 2004. Variations among somaclones and its seedling progeny in Capsicum annuum. Plant Cell Tissue Org. Cult. 76:261-267. 2. Anu, A. and K.V. Peter. 2003. Analysis of seed protein of 29 lines of Capsicum annuum L. by polyacrylamide gel electrophoresis. Genet. Resour. Crop Evol. 50:239-243. 3. Dwivedi, N., R. Kumar, R. Paliwal, R. Kumar, S. Kumar, M. Singh and R.K. Singh. 2013. QTL mapping for important horticultural traits in pepper (Capsicum annuum L.). J. Plant Bioch. Biotech. doi: 10.1007/s13562-013-0247-1. 4. FAOSTAT. 2012. <http://faostat.fao.org/site/339/default.aspx> 5. Garg, R., S. Kumar, R. Kumar, M. Loganathan, S. Saha, S. Kumar, A.B. Rai, and B.K. Roy. 2013. Novel source of resistance and differential reactions on chilli fruit infected by Colletotrichum capsici. Australasian Plant Pathol. 42:227-233. 6. Kumar, S., V. Singh, M. Singh, S. Rai, S. Kumar, S.K. Rai, and M. Rai. 2007. Genetics and validation of fertility restoration associated RAPD markers in pepper (Capsicum annuum L.). Scientia Hort. 111:197-202. 7. Kumar, R., S. Kumar, N. Dwivedi, S. Kumar, A. Rai, M. Singh, D.S. Yadav, and M. Rai. 2009. Validation of SCAR markers, diversity analysis of male sterile cytoplasms (S-) and isolation of an alloplasmic S-cytoplasm in Capsicum. Scientia Hort. 120:167-172. 8. Kumar, S., R. Kumar, S. Kumar, M. Singh, A.B. Rai, and M. Rai. 2011. Incidences of leaf curl disease on Capsicum germplasm under field conditions. Indian J. Agr. Sci. 81:187-189. 9. Kumar, R.V., V.K. Sharma, B. Chattopadhyay, and S. Chakraborty. 2012. An improved plant regeneration and Agrobacterium mediated transformation of red pepper (Capsicum annuum L.). Physiol. Mol. Biol. Plants 18:357-364. 10. Kumar, S., S. Kumar, H.C. Shieh, S.W. Lin, R. Schafleitner, L. Kenyon, R. Srinivasan, J.F. Wang, A.W. Ebert, and Y.Y. Chou. 2014. Domestication of peppers (Capsicum spp.) and breeding for global use. Second Intl. Plant Breed. Seminar, Universiti Putra Malaysia (UPM) (14-15 October), Serdang, Malaysia. 11. Lin, S.W., Y.Y. Chou, H.C. Shieh, A.W. Ebert, S. Kumar, R. Mavlyanova, A. Rouamba, A. Tenkouano, V. Afari-Sefa, and P.A. Gniffke. 2013. Pepper (Capsicum spp.) germplasm dissemination by AVRDC The World Vegetable Center: an overview and introspection. Chronica Hort. 53:21-27. 12. Rai, V.P., R. Kumar, S. Kumar, A. Rai, A. Kumar, M. Singh, S.P. Singh, A.B. Rai, and R. Paliwal. 2013. Genetic diversity in Capsicum germplasm based on microsatellite and random amplified microsatellite polymorphism markers. Physiol. Mol. Biol. Plant 19:575-586.

Exploitation of AVRDC s Chili Pepper (Capsicum spp.) Germplasm in India -9 13. Rai, V.P., R. Kumar, S.P. Singh, S. Kumar, S. Kumar, M. Singh, and M. Rai. 2014. Monogenic recessive resistance to Pepper leaf curl virus in an interspecific cross of Capsicum. Scientia Hort.172:34-38. 14. Reddy, M.K., A. Srivastava, S. Kumar, R. Kumar, N. Chawda, A.W. Ebert, and M. Vishwakarma. 2014. Chilli (Capsicum annuum L.) breeding in India: An overview. SABRAO J. Breed. Genet. 46:160-173. 15. Reddy, K.M. and M.K. Reddy. 2010. Breeding for virus resistance. p. 119-132. In: R. Kumar, A.B. Rai, M. Rai, and H.P. Singh (eds.), Advances in chilli research, Studium Press Pvt. Ltd., New Delhi, India. 16. Singh, P., D.S. Cheema, M.S. Dhaliwal, and N. Garg. 2014. Heterosis and combining ability for earliness, plant growth, yield and fruit attributes in hot pepper (Capsicum annuum L.) involving genetic and cytoplasmic-genetic male sterile lines. Scientia Hort. 168:175-188. 摘要 辣椒對印度的農民為很重要蔬果作物, 番椒 ( 辣椒與甜椒 ) 在印度是種植面積最廣的作物 在 2012-13 年間, 印度出口辣椒以及其相關製品產值約有 4.1 億美金 在過去的 13 年中, 亞蔬 - 世界蔬菜中心已提供超過 33,000 個番椒試驗種原, 包括改良品系 ( 佔 81%) 和種原 (19%) 至世界各國 120 多個國家的公 私立單位, 印度居最高種原申請國, 共 5187 個番椒種子樣本, 佔全球申請總樣本的 15.6%, 這些種原和改良自交品系成功被印度利用成為商業自交品種或雜交一代品種 在 2012-2013 年間, 估計這些由亞蔬 - 世界蔬菜中心的種原和改良品系所育成的雜交一代品種, 已在印度的各個不同地區種植, 面積超過 30,000 ha

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