中国作物种质资源安全保存理论与实践
|
摘要
我国是作物种质资源大国,目前已经收集保存了340种作物,保存资源总量达50万份,如何确保安全保存是种质资源管理的首要课题。自"九五"以来,系统地开展了作物种质资源安全保存理论与技术的研究,揭示了种质活力丧失存在关键节点及其生物学机制;明确了发芽率低于关键节点后进行更新,会导致群体遗传完整性降低;提出了种质安全保存寿命的概念,以及延长安全保存寿命的关键因素。研发了种质活力监测预警、繁殖更新与离体保存等关键技术,制定了种质资源入库圃保存、监测预警、繁殖更新、离体保存和设施建设等技术规程,创建了中国作物种质资源安全保存技术体系。该体系应用于全国作物种质资源的保存实践,最大程度延长种质安全保存寿命,并监测预警出需更新的种质,避免因活力和遗传完整性丧失而导致种质资源得而复失,为实现我国作物种质资源长久安全保存提供了有力保障。
China has abundant crop germplasm resources,and has preserved over 500000 accessions of 340 crops.Ensuring the safe conservation of germplasm resources is therefore of high priority for genebank managers.Since the 9th Five-Year Plan,the National Crop Genebank of China has systematically studied the theories and technologies of the safe conservation of germplasm.For the first time,we proposed a theory for germplasm resources conservation,including both safety backup and the biological mechanism of conservation.The transition from platform(high viability)to the rapid reduction phase of the seed survival curve was defined as the critical node(CN).It was revealed that oxidative damage and mitochondrial damage disordered the metabolism during the CN.In addition,germplasm would loss genetic integrity,if regenerated when its germination rate was lower than the CN.Taking loss of viability and genetic integrity together,germplasm longevity is defined as the time taking by seed viability declined to the CN.We have made breakthroughs on the key techniques for seed viability monitoring,early warning of viability decline,regeneration,in vitro conservation and cryopreservation.Based on the above research,we have created systematic technologies for the safe conservation of crop germplasm resources,which have been applied national-widely.To avoid the loss of viability and genetic integrity in genebank,the systematic technologies would prolong the germplasm longevity,monitor and forecast seed viability,recognize the germplasm which needs regeneration.Therefore,the systematic technologies provide reliable guarantee for the safe conservation of crop germplasm resources.
China has abundant crop germplasm resources,and has preserved over 500000 accessions of 340 crops.Ensuring the safe conservation of germplasm resources is therefore of high priority for genebank managers.Since the 9th Five-Year Plan,the National Crop Genebank of China has systematically studied the theories and technologies of the safe conservation of germplasm.For the first time,we proposed a theory for germplasm resources conservation,including both safety backup and the biological mechanism of conservation.The transition from platform(high viability)to the rapid reduction phase of the seed survival curve was defined as the critical node(CN).It was revealed that oxidative damage and mitochondrial damage disordered the metabolism during the CN.In addition,germplasm would loss genetic integrity,if regenerated when its germination rate was lower than the CN.Taking loss of viability and genetic integrity together,germplasm longevity is defined as the time taking by seed viability declined to the CN.We have made breakthroughs on the key techniques for seed viability monitoring,early warning of viability decline,regeneration,in vitro conservation and cryopreservation.Based on the above research,we have created systematic technologies for the safe conservation of crop germplasm resources,which have been applied national-widely.To avoid the loss of viability and genetic integrity in genebank,the systematic technologies would prolong the germplasm longevity,monitor and forecast seed viability,recognize the germplasm which needs regeneration.Therefore,the systematic technologies provide reliable guarantee for the safe conservation of crop germplasm resources.
引文
[1]FAO.Second report on the state of the world's plants genetic resources for food and agriculture.Commission on Genetic Resources for Food and Agriculture,Rome,2010,47
[2]Walters C,Wheeler L M,Grotenhuis J M.Longevity of seeds stored in a genebank:species characteristics.Seed Science Research,2005,15:1-20
[3]卢新雄,崔聪淑,陈晓玲,陈贞,陈叔平.国家种质库部分作物种子生活力监测结果与分析.植物遗传资源科学,2001,2(2):1-5Lu X X,Cui C S,Chen X L,Chen Z,Chen S P.Survey of seed germinability after 10~12 years storage in the National Genebank of China.Journal of Plant Genetic Resources,2001,2(2):1-5
[4]辛霞,陈晓玲,张金梅,卢新雄.国家库贮藏20年以上种子生活力与田间出苗率监测.植物遗传资源学报,2011,12(6):934-940Xin X,Chen X L,Zhang J M,Lu X X.Germinability and seedling emergence of seeds after 20 years of storage in the National Genebank of China.Journal of Plant Genetic Resources,2011,12(6):934-940
[5]Stoyanova S D.Effect of seed ageing and regeneration on the genetic composition of wheat.Seed Science Technology,1992,20:489-496
[6]Chwedorzewska K J,Bednarek P T,Puchalski J,Krajewski P.AFLP-profiling of long-term stored and regenerated rye genebank samples.Cellular and Molecular Biology Letters,2002,7:457-463
[7]Soengas P,Cartea E,Lema M,Velasco P.Effect of regeneration procedures on the genetic integrity of Brassica oleracea accessions.Molecular Breeding,2009,23:389-395
[8]Dulloo M E,Hanson J,Jorge M A,Thormann I.Regeneration guidelines:general guiding principles,2008
[9]杨勇,赵红星,李高潮,阮小凤,王仁梓.柿种质资源田间耐寒性调查分析.北方园艺,2010(1):63-65Yang Y,Zhao H X,Li G C,Ruan X F,Wang R Z.Field investigation of cold tolerance in persimmon(Diospyros L.)germplasm.Northern Horticulture,2010(1):63-65
[10]卢新雄,陈晓玲,陈叔平.低温库种质安全保存理论的研究进展.植物遗传资源科学,2000,1(2):54-58Lu X X,Chen X L,Chen S P.Advances on theory of the safe conservaton for germplasm in seed Genebank.Journal of Plant Genetic Resources,2000,1(2):54-58
[11]卢新雄,崔聪淑,陈晓玲,张晗,李秀全,李高原,陈丽华.小麦种质贮藏过程中生活力丧失特性及田间出苗率表现.植物遗传资源学报,2003,4(3):220-224Lu X X,Cui C S,Chen X L,Zhang H,Li X Q,Li G Y,Chen L H.Characteristics of wheat seed viability loss during storage and emergence rate of seedlings.Journal of Plant Genetic Resources,2003,4(3):220-224
[12]卢新雄,陈晓玲.水稻种子贮藏过程中生活力丧失特性及预警指标的研究.中国农业科学,2002,35(8):975-979Lu X X,Chen X L.Characteristics and warning indices of rice seeds viability loss during storage at 45℃constant temperature.Scientia Agricultura Sinica,2002,35(8):975-979
[13]辛霞,陈晓玲,张金梅,卢新雄.小麦种子在不同保存条件下的生活力丧失特性研究.植物遗传资源学报,2013,14(4):588-593Xin X,Chen X L,Zhang J M,Lu X X.Viability loss of wheat seeds under different storage conditions.Journal of Plant Genetic Resources,2013,14(4):588-593
[14]Yin G K,Whelan J,Wu S H,Zhou J,Chen B Y,Chen X L,Zhang J M,He J J,Xin X,Lu X X.Comprehensive mitochondrial metabolic shift during the critical node of seed ageing in rice.PLoS ONE,2016,11(4):e0148013
[15]Yin G K,Xin X,Fu S Z,An M N,Wu S H,Chen X L,Zhang J M,He J J,Whelan J,Lu X X.Proteomic and carbonylation profile analysis at the critical node of seed ageing in Oryza sativa.Scientific Reports,2017,7:1-12
[16]FAO.Genebank Standards.Food and Agriculture Organization of The United Nations,Rome,2014,36
[17]Xin X,Lin X H,Zhou Y C,Chen X L,Liu X,Lu X X.Proteome analysis of maize seeds:the effect of artificial ageing.Physiologia Plantarum,2011,143(2):126-138
[18]Yin G K,Xin X,Song C,Chen X L,Zhang J M,Wu S H,Li RF,Liu X,Lu X X.Activity levels and expression of antioxidant enzymes in the ascorbate-glutathione cycle in artificially aged rice seed.Plant Physiology and Biochemistry,2014,80:1-9
[19]Chen X L,Yin G K,B?rner A,Xin X,He J J,Nagel M,Liu X,Lu X X.Comparative physiology and proteomics of two wheat genotypes differing in seed storage tolerance.Plant Physiology and Biochemistry,2018,130:455-463
[20]Fu S Z,Yin G K,Xin X,Wu S H,Wei X H,Lu X X.The levels of crotonaldehyde and 4-hydroxy-(E)-2-nonenal and carbonylscavenging enzyme gene expression at the critical node during rice seed aging.Rice Science,2018,25(3):152-160
[21]Xin X,Tian Q,Yin G K,Chen X L,Zhang J M,Ng S,Lu X X.Reduced mitochondrial and ascorbate-glutathione activity after artificial ageing in soybean seed.Journal of Plant Physiology,2014,171(2):140-147
[22]Kühn K,Yin G K,Duncan O,Law S R,Kubiszewski-Jakubiak S,Kaur P,Meyer E,Wang Y,des Francs Small C C,Giraud E,Narsai R,Whelan J.Decreasing electron flux through the cytochrome and/or alternative respiratory pathways triggers common and distinct cellular responses dependent on growth conditions.Plant Physiology,2015,167(1):228-250
[23]Van Hintum T J Th,van de Wiel C C M,visser D L,van Treuren R,Vosman B.The distribution of genetic diversity in a Brassica oleracea gene bank collection related to the effects on diversity of regeneration,as measured with AFLPs.Theoretical and Applied Genetics,2007,114:777-786
[24]张晗,卢新雄,张志娥,陈晓玲,任守杰,辛萍萍.种子老化对玉米种质资源遗传完整性变化的影响.植物遗传资源学报,2005,6(3):271-275Zhang H,Lu X X,Zhang Z E,Chen X L,Ren S J,Xin PP.Effect of seed aging on changes of genetic integrity in maize germplasm.Journal of Plant Genetic Resources,2005,6(3):271-275
[25]王栋,卢新雄,张志娥,陈晓玲,辛霞,辛萍萍,耿立格.SSR标记分析种子老化及繁殖世代对大豆种质遗传完整性的影响.植物遗传资源学报,2010,11(2):192-199Wang D,Lu X X,Zhang Z E,Chen X L,Xin X,Xin P P,Geng L G.Effect of seed aging and regeneration on genetic integrity in soybean by SSR markers.Journal of Plant Genetic Resources,2010,11(2):192-199
[26]王栋,张志娥,陈晓玲,辛霞,辛萍萍,卢新雄.AFLP标记分析生活力影响大豆中黄18种质遗传完整性.作物学报,2010,36(4):555-564Wang D,Zhang Z E,Chen X L,Xin X,Xin P P,Lu X X.Analysis of viability affecting on genetic integrity in soybean germplasm Zhonghuang 18 by AFLP markers.Acta Agronomica Sinica,2010,36(4):555-564
[27]刘敏,辛霞,张志娥,陈晓玲,张金梅,卢新雄.繁殖群体量及隔离对蚕豆种质遗传完整性的影响.植物遗传资源学报,2012,13(2):175-181Liu M,Xin X,Zhang Z E,Chen X L,Zhang J M,Lu X X.Effect of sample sizes and isolation methods on genetic integrity of faba bean(Vicia faba L.)Germplasm.Journal of Plant Genetic Resources,2012,13(2):175-181
[28]宋超,辛霞,陈晓玲,张金梅,尹广鹍,何娟娟,覃初贤,卢新雄.三种保存条件下水稻和小麦种质资源安全保存期的分析.植物遗传资源学报,2014,15(4):685-691Song C,Xin X,Chen X L,Zhang J M,Yin G K,He J J,Qin C X,Lu X X.Safety storage life of rice and wheat germplasm resources at three storage conditions.Journal of Plant Genetic Resources,2014,15(4):685-691
[29]陈贞.大豆种子生活力跟踪监测研究.种子,1999,4:46-48Chen Z.Study on tracking and monitoring of soybean seed viability.Seed,1999,4:46-48
[30]卢新雄,崔聪淑,陈晓玲,陈贞,陈叔平.国家种质库部分作物种子生活力监测结果与分析.植物遗传资源科学,2001,2(2):1-5Lu X X,Cui C S,Chen X L,Chen Z,Chen S P.Survey of seed germinability after 10~12 years storage in the National Genebank of China.Journal of Plant Genetic Resources,2001,2(2):1-5
[31]胡群文,卢新雄,辛萍萍,陈晓玲,张志娥,辛霞,刘旭.水稻在不同气候区室温贮藏的适宜含水量及存活特性.中国水稻科学,2009,23(6):621-627Hu Q W,Lu X X,Xin P P,Chen X L,Zhang Z E,Xin X,Liu X.The optimal moisture content and survival characteristics of rice seeds stored at six climatic zones under room temperature.Chinese Journal of Rice Science,2009,23(6):621-627
[32]卢新雄,张云兰.国家种质库种子干燥处理技术的建立与应用.植物遗传资源学报,2003,4(4):365-368Lu X X,Zhang Y L.Establishment and use of seed drying technology in the National Crop Genebank of China.Journal of Plant Genetic Resources,2003,4(4):365-368
[33]吴晓亮,辛萍萍,张志娥,陈晓玲,陶澜,卢新雄.水稻种子室温贮藏最适含水量及其热稳定蛋白的研究.中国农业科学,2006,39(11):2214-2219Wu X L,Xin P P,Zhang Z E,Chen X L,Tao L,Lu X X.Studies on optimum moisture content and heat-stable protein of rice seeds stored at room temperature.Scientia Agricultura Sinica,2006,39(11):2214-2219
[34]周静,辛霞,尹广鹍,陈晓玲,张金梅,卢新雄.大豆种子在不同气候区室温贮藏的适宜含水量与寿命关系研究.大豆科学,2014,33(5):687-690Zhou J,Xin X,Yin G K,Chen X L,Zhang J M,Lu X X.Optimal moisture content and longevity of soybean seeds stored at different climatic zones under ambient condition.Soybean Science,2014,33(5):687-690
[35]辛霞,张金梅,尹广鹍,赵婧,陈晓玲,卢新雄.快速无损检测种子生活力的方法.ZL201510847974.8.2018-03-23Xin X,Zhang J M,Yin G K,Zhao J,Chen X L,Lu XX.A fast,non-invasive technique for seed viability testing.ZL201510847974.8.2018-03-23
[36]辛霞,赵婧,陈晓玲,尹广鹍,张金梅,卢新雄.快速无损检测大豆种子生活力的方法.ZL201510849232.9.2018-02-02Xin X,Zhao J,Chen X L,Yin G K,Zhang J M,Lu X X.Afast,non-invasive technique for soybean seed viability testing.ZL201510849232.9.2018-02-02
[37]辛霞,尹广鹍,陈晓玲,张金梅,赵婧,卢新雄.快速无损检测小麦种子生活力的方法.ZL201510849640.4.2018-08-17Xin X,Yin G K,Chen X L,Zhang J M,Zhao J,Lu X X.Afast,non-invasive technique for wheat seed viability testing.ZL201510849640.4.2018-08-17
[38]Xin X,Wan Y L,Wang W J,Yin G K,ES M L,Lu X X.Areal-time,non-invasive,micro-optrode technique for detecting seed viability by using oxygen influx.Scientific Reports,2013,3(10):3507
[39]王述民,卢新雄,李立会.作物种质资源繁殖更新技术规程.北京:中国农业科学技术出版社,2014Wang S M,Lu X X,Li L H.The technical regulation on regeneration of crop germplasm resources.Beijing:China Agricultural Science and Technology Press,2014
[40]Benson E.E.Cryopreservation of phytodiversity:A critical appraisal of theory practice.Critical Reviews in Plant Sciences,2008,27(3):141-219
[41]Engelmann F.Use of biotechnologies for the conservation of plant biodiversity.In Vitro Cellular and Developmental BiologyPlant,2011,47(1):5-16
[42]Panis B,Swennen R,Rhee J,Roux N.Securing plant genetic resources for perpetuity through cryopreservation.Indian Journal of Plant Genetic Resources,2016,29(3):300-302
[43]International Potato Center.Number of accessions in the CIPCryobank.(2018-08-12)www.cipotato.org/genebankcip/process/cryopreservation-2
[44]Jenderek M M,Reed B M.Cryopreserved storage of clonal germplasm in the USDA National Plant Germplasm System.In Vitro Cellular and Developmental Biology-Plant,2017,53(4):299-308
[45]陈辉,陈晓玲,陈龙清,卢新雄.切花百合离体茎尖玻璃化法超低温保存研究.植物遗传资源学报,2007,8(2):170-173Chen H,Chen X L,Chen L Q,Lu X X.Cryopreservation of shoot tips from in vitro plants of cut flower of lily(Lilium L.)by vitrification method.Journal of Plant Genetic Resources,2007,8(2):170-173
[46]赵艳华,吴雅琴,程和禾,李春敏,陈晓玲,卢新雄.李离体茎尖的超低温保存.园艺学报,2008,35(3):423-426Zhao Y H,Wu Y Q,Cheng H M,Li C M,Chen X L,Lu XX.Cryopreservation of shoot tips from prunus.Acta Horticulturae Sinica,2008,35(3):423-426
[47]刘丽芳.甘薯种质遗传稳定性及超低温保存研究.重庆:西南大学,2009Liu L F.Studies on Genetic Stability and Cryopreservation of Sweet Potato Germplasm.Chongqing:Southwest University,2009
[48]白建明,陈晓玲,卢新雄,郭华春,辛霞,张志娥,辛萍萍.马铃薯茎尖小滴玻璃化法超低温保存及其再生植株的遗传稳定性.园艺学报,2010,37(9):1431-1438Bai J M,Chen X L,Lu X X,Guo H C,Xin X,Zhang Z E,Xin P P.Cryopreservation of in vitro shoot tips of potato by droplet vitrification and genetic stability of regenerated plantlets.Acta Horticulturae Sinica,2010,37(9):1431-1438
[49]李俊慧,何平,陈晓玲,卢新雄,辛霞,张志娥,辛萍萍.香蕉离体茎尖超低温保存研究.植物遗传资源学报,2010,11(1):34-39Li J H,He P,Chen X L,Lu X X,Xin X,Zhang Z E,Xin P P.Cryopreservation of in vitro shoot-tips of banana(Musa spp.).Journal of Plant Genetic Resources,2010,11(1):34-39
[50]Chen X L,Li J H,Xin X,Zhang Z E,Xin P P,Lu X X.Cryopreservation of in vitro-grown apical meristems of Lilium by dropletvitrification.South African Journal of Botany,2011,77(2):397-403
[51]Zhang J M,Zhang X N,Lu X X,Xin X,Yin G K,He J J,Xu Y M,Chen X L.Optimization of droplet-vitrification protocol for carnation genotypes and ultra-structural studies on shoot tips during cryopreservation.Acta Physiologiae Plantarum,2014,36(12):3189-3198
[52]Zhang J M,Huang B,Zhang X N,Volk G M,Zhou Y C,Chen X L.Identification of a highly successful cryopreservation method(droplet-vitrification)for petunia.In Vitro Cellular and Developmental Biology-Plant,2015,51(4):445-451
[53]Zhang J M,Huang B,Lu X X,Volk G M,Xin X,Yin G K,He J J,Chen X L.Cryopreservation of in vitro-grown shoot tips of Chinese medicinal plant Atractylodes macrocephala Koidz.using a droplet-vitrification method.CryoLetters,2015,36(3):195-204
[54]Zhang J M,Han L,Lu X X,Volk G M,Xin X,Yin G K,He JJ,Wang L,Chen X L.Cryopreservation of Jerusalem artichoke cultivars using an improved droplet-vitrification method.Plant Cell and Tissue Organ Culture,2017,128(3):577-587
[55]Zhang J M,Lu X X,Xin X,Yin G K,He J J,Huang B,Jiang D,Chen X L.Cryopreservation of Citrus anthers in the National Crop Genebank of China.In Vitro Cellular and Developmental Biology-Plant,2017,51(4):445-451
[56]张金梅,闫文君,李雪,黄斌,张海晶,赵剑波,姜全,陈晓玲.桃花粉低温和超低温保存方法比较研究.植物遗传资源学报,2017,18(4):670-675Zhang J M,Yan W J,Li X,Huang B,Zhang H J,Zhao J B,Jiang Q,Chen X L.Study on low temperature storage and cryopreservation of pollen for peach germplasm resources.Journal of Plant Genetic Resources,2017,18(4):670-675
[57]陈晓玲,张金梅,辛霞,卢新雄.山葵试管苗离体茎尖包埋干燥超低温保存及再生培养方法,ZL201210211725.6.2014-04-02Chen X L,Zhang J M,Xin X,Lu X X.Methods of cryopreservation and regeneration for in vitro-grown shoot tips of wasabi.ZL201210211725.6.2014-04-02
[58]陈晓玲,张金梅,卢新雄,辛霞,尹广鹍,何娟娟.桑树休眠芽段的超低温保存及嫁接复活方法,ZL201410366753.4.2016-06-11Chen X L,Zhang J M,Lu X X,Xin X,Yin G K,He J J.Methods of cryopreservation and grafting regeneration for mulberry dormant bud.ZL201410366753.4.2016-06-11
[59]Huang B,Zhang J M,Chen X L,Xin X,Yin G K,He J J,Lu X X,Zhou Y C.Oxidative damage and antioxidative indicators in 48 h germinated rice embryos during the vitrificationcryopreservation procedure.Plant Cell Reports,2018,37:1325-1342
[60]李运合,李玉生,张金梅,陈晓玲,赵艳华,吴永杰.芒果细胞超低温保存存活机理.植物生理学报,2016,52(10):1474-1480Li Y H,Li Y S,Zhang J M,Chen X L,Zhao Y H,Wu Y J.The survival mechanism of mango cells during cryopreservation.Plant Physiology Journal,2016,52(10):1474-1480
[61]张海晶,张金梅,辛霞,尹广鹍,何娟娟,卢新雄,周元昌,陈晓玲.马铃薯茎尖超低温保存流程TTC活力响应.植物遗传资源学报,2015,16(3):555-560Zhang H J,Zhang J M,Xin X,Yin G K,He J J,Lu X X,Zhou Y C,Chen X L.Study on potato shoot tips vitality responses during cryopreservation by TTC staining method.Journal of Plant Genetic Resources,2015,16(3):555-560
[62]Zhang J M,Xin X,Yin G K,Lu X X,Chen X L.In vitro conservation and cryopreservation in National Genebank of China.Acta Horticulturae,2014,1039(1039):309-317
[63]陈晓玲,张金梅,辛霞,黄斌,卢新雄.植物种质资源超低温保存现状及其研究进展.植物遗传资源学报,2013,14(3):414-427Chen X L,Zhang J M,Xin X,Huang B,Lu X X.Progress on cryopreservation state and research of plant germplasm resources.Journal of Plant Genetic Resources,2013,14(3):414-427
[64]何娟娟,卢新雄,栗钊,辛霞.8种粮食和蔬菜作物种子短期节能储藏研究.中国种业,2016(9):57-60He J J,Lu X X,Li Z,Xin X.Short-term storage of 8 crop and vegetable seeds with an energy-saving method.China Seed Industry,2016(9):57-60
[65]卢新雄,陈叔平,刘旭.农作物种质资源保存技术规程.北京:中国农业出版社,2008Lu X X,Chen S P,Liu X.The technical regulation on conservation of crop germplasm resources,Beijing:China Agriculture Press,2008
[66]卢新雄.农业种质库的设计与建设要求探讨.农业工程学报,2003,19(6):252-255Lu X X.Design and construction of agricultural germplasm genebank.Transactions of The Chinese Society of Agricultural Engineering,2003,19(6):252-255
[67]郭颖荷,卢新雄,彭高军.中国低温种质库建造技术调查分析.农业工程学报,2005,21(4):186-190Guo Y H,Lu X X,Peng G J.Investigation and analysis of the building technology of low temperature germplasm genebanks of China.Transactions of The Chinese Society of Agricultural Engineering,2005,21(4):186-190
[68]中华人民共和国农业农村部,中华人民共和国国家发展和改革委员会,中华人民共和国科学技术部.全国农作物种质资源保护与利用中长期发展规划(2015-2030年).(2015-04-29)[2018-08-15]http://www.moa.gov.cn/nybgb/2015/si/201711/t20171129_6134098.htm Ministry of Agriculture of the People's Republic of China,National Development and Reform Commission of the People's Republic of China,People's Republic of China Ministry of Science and Technology.National Medium-and Long-term Development Plan for the Protection and Utilization of Crop Germplasm Resources(2015-2030).(2015-04-29)[2018-08-15]http://www.moa.gov.cn/nybgb/2015/si/201711/t20171129_6134098.htm
[2]Walters C,Wheeler L M,Grotenhuis J M.Longevity of seeds stored in a genebank:species characteristics.Seed Science Research,2005,15:1-20
[3]卢新雄,崔聪淑,陈晓玲,陈贞,陈叔平.国家种质库部分作物种子生活力监测结果与分析.植物遗传资源科学,2001,2(2):1-5Lu X X,Cui C S,Chen X L,Chen Z,Chen S P.Survey of seed germinability after 10~12 years storage in the National Genebank of China.Journal of Plant Genetic Resources,2001,2(2):1-5
[4]辛霞,陈晓玲,张金梅,卢新雄.国家库贮藏20年以上种子生活力与田间出苗率监测.植物遗传资源学报,2011,12(6):934-940Xin X,Chen X L,Zhang J M,Lu X X.Germinability and seedling emergence of seeds after 20 years of storage in the National Genebank of China.Journal of Plant Genetic Resources,2011,12(6):934-940
[5]Stoyanova S D.Effect of seed ageing and regeneration on the genetic composition of wheat.Seed Science Technology,1992,20:489-496
[6]Chwedorzewska K J,Bednarek P T,Puchalski J,Krajewski P.AFLP-profiling of long-term stored and regenerated rye genebank samples.Cellular and Molecular Biology Letters,2002,7:457-463
[7]Soengas P,Cartea E,Lema M,Velasco P.Effect of regeneration procedures on the genetic integrity of Brassica oleracea accessions.Molecular Breeding,2009,23:389-395
[8]Dulloo M E,Hanson J,Jorge M A,Thormann I.Regeneration guidelines:general guiding principles,2008
[9]杨勇,赵红星,李高潮,阮小凤,王仁梓.柿种质资源田间耐寒性调查分析.北方园艺,2010(1):63-65Yang Y,Zhao H X,Li G C,Ruan X F,Wang R Z.Field investigation of cold tolerance in persimmon(Diospyros L.)germplasm.Northern Horticulture,2010(1):63-65
[10]卢新雄,陈晓玲,陈叔平.低温库种质安全保存理论的研究进展.植物遗传资源科学,2000,1(2):54-58Lu X X,Chen X L,Chen S P.Advances on theory of the safe conservaton for germplasm in seed Genebank.Journal of Plant Genetic Resources,2000,1(2):54-58
[11]卢新雄,崔聪淑,陈晓玲,张晗,李秀全,李高原,陈丽华.小麦种质贮藏过程中生活力丧失特性及田间出苗率表现.植物遗传资源学报,2003,4(3):220-224Lu X X,Cui C S,Chen X L,Zhang H,Li X Q,Li G Y,Chen L H.Characteristics of wheat seed viability loss during storage and emergence rate of seedlings.Journal of Plant Genetic Resources,2003,4(3):220-224
[12]卢新雄,陈晓玲.水稻种子贮藏过程中生活力丧失特性及预警指标的研究.中国农业科学,2002,35(8):975-979Lu X X,Chen X L.Characteristics and warning indices of rice seeds viability loss during storage at 45℃constant temperature.Scientia Agricultura Sinica,2002,35(8):975-979
[13]辛霞,陈晓玲,张金梅,卢新雄.小麦种子在不同保存条件下的生活力丧失特性研究.植物遗传资源学报,2013,14(4):588-593Xin X,Chen X L,Zhang J M,Lu X X.Viability loss of wheat seeds under different storage conditions.Journal of Plant Genetic Resources,2013,14(4):588-593
[14]Yin G K,Whelan J,Wu S H,Zhou J,Chen B Y,Chen X L,Zhang J M,He J J,Xin X,Lu X X.Comprehensive mitochondrial metabolic shift during the critical node of seed ageing in rice.PLoS ONE,2016,11(4):e0148013
[15]Yin G K,Xin X,Fu S Z,An M N,Wu S H,Chen X L,Zhang J M,He J J,Whelan J,Lu X X.Proteomic and carbonylation profile analysis at the critical node of seed ageing in Oryza sativa.Scientific Reports,2017,7:1-12
[16]FAO.Genebank Standards.Food and Agriculture Organization of The United Nations,Rome,2014,36
[17]Xin X,Lin X H,Zhou Y C,Chen X L,Liu X,Lu X X.Proteome analysis of maize seeds:the effect of artificial ageing.Physiologia Plantarum,2011,143(2):126-138
[18]Yin G K,Xin X,Song C,Chen X L,Zhang J M,Wu S H,Li RF,Liu X,Lu X X.Activity levels and expression of antioxidant enzymes in the ascorbate-glutathione cycle in artificially aged rice seed.Plant Physiology and Biochemistry,2014,80:1-9
[19]Chen X L,Yin G K,B?rner A,Xin X,He J J,Nagel M,Liu X,Lu X X.Comparative physiology and proteomics of two wheat genotypes differing in seed storage tolerance.Plant Physiology and Biochemistry,2018,130:455-463
[20]Fu S Z,Yin G K,Xin X,Wu S H,Wei X H,Lu X X.The levels of crotonaldehyde and 4-hydroxy-(E)-2-nonenal and carbonylscavenging enzyme gene expression at the critical node during rice seed aging.Rice Science,2018,25(3):152-160
[21]Xin X,Tian Q,Yin G K,Chen X L,Zhang J M,Ng S,Lu X X.Reduced mitochondrial and ascorbate-glutathione activity after artificial ageing in soybean seed.Journal of Plant Physiology,2014,171(2):140-147
[22]Kühn K,Yin G K,Duncan O,Law S R,Kubiszewski-Jakubiak S,Kaur P,Meyer E,Wang Y,des Francs Small C C,Giraud E,Narsai R,Whelan J.Decreasing electron flux through the cytochrome and/or alternative respiratory pathways triggers common and distinct cellular responses dependent on growth conditions.Plant Physiology,2015,167(1):228-250
[23]Van Hintum T J Th,van de Wiel C C M,visser D L,van Treuren R,Vosman B.The distribution of genetic diversity in a Brassica oleracea gene bank collection related to the effects on diversity of regeneration,as measured with AFLPs.Theoretical and Applied Genetics,2007,114:777-786
[24]张晗,卢新雄,张志娥,陈晓玲,任守杰,辛萍萍.种子老化对玉米种质资源遗传完整性变化的影响.植物遗传资源学报,2005,6(3):271-275Zhang H,Lu X X,Zhang Z E,Chen X L,Ren S J,Xin PP.Effect of seed aging on changes of genetic integrity in maize germplasm.Journal of Plant Genetic Resources,2005,6(3):271-275
[25]王栋,卢新雄,张志娥,陈晓玲,辛霞,辛萍萍,耿立格.SSR标记分析种子老化及繁殖世代对大豆种质遗传完整性的影响.植物遗传资源学报,2010,11(2):192-199Wang D,Lu X X,Zhang Z E,Chen X L,Xin X,Xin P P,Geng L G.Effect of seed aging and regeneration on genetic integrity in soybean by SSR markers.Journal of Plant Genetic Resources,2010,11(2):192-199
[26]王栋,张志娥,陈晓玲,辛霞,辛萍萍,卢新雄.AFLP标记分析生活力影响大豆中黄18种质遗传完整性.作物学报,2010,36(4):555-564Wang D,Zhang Z E,Chen X L,Xin X,Xin P P,Lu X X.Analysis of viability affecting on genetic integrity in soybean germplasm Zhonghuang 18 by AFLP markers.Acta Agronomica Sinica,2010,36(4):555-564
[27]刘敏,辛霞,张志娥,陈晓玲,张金梅,卢新雄.繁殖群体量及隔离对蚕豆种质遗传完整性的影响.植物遗传资源学报,2012,13(2):175-181Liu M,Xin X,Zhang Z E,Chen X L,Zhang J M,Lu X X.Effect of sample sizes and isolation methods on genetic integrity of faba bean(Vicia faba L.)Germplasm.Journal of Plant Genetic Resources,2012,13(2):175-181
[28]宋超,辛霞,陈晓玲,张金梅,尹广鹍,何娟娟,覃初贤,卢新雄.三种保存条件下水稻和小麦种质资源安全保存期的分析.植物遗传资源学报,2014,15(4):685-691Song C,Xin X,Chen X L,Zhang J M,Yin G K,He J J,Qin C X,Lu X X.Safety storage life of rice and wheat germplasm resources at three storage conditions.Journal of Plant Genetic Resources,2014,15(4):685-691
[29]陈贞.大豆种子生活力跟踪监测研究.种子,1999,4:46-48Chen Z.Study on tracking and monitoring of soybean seed viability.Seed,1999,4:46-48
[30]卢新雄,崔聪淑,陈晓玲,陈贞,陈叔平.国家种质库部分作物种子生活力监测结果与分析.植物遗传资源科学,2001,2(2):1-5Lu X X,Cui C S,Chen X L,Chen Z,Chen S P.Survey of seed germinability after 10~12 years storage in the National Genebank of China.Journal of Plant Genetic Resources,2001,2(2):1-5
[31]胡群文,卢新雄,辛萍萍,陈晓玲,张志娥,辛霞,刘旭.水稻在不同气候区室温贮藏的适宜含水量及存活特性.中国水稻科学,2009,23(6):621-627Hu Q W,Lu X X,Xin P P,Chen X L,Zhang Z E,Xin X,Liu X.The optimal moisture content and survival characteristics of rice seeds stored at six climatic zones under room temperature.Chinese Journal of Rice Science,2009,23(6):621-627
[32]卢新雄,张云兰.国家种质库种子干燥处理技术的建立与应用.植物遗传资源学报,2003,4(4):365-368Lu X X,Zhang Y L.Establishment and use of seed drying technology in the National Crop Genebank of China.Journal of Plant Genetic Resources,2003,4(4):365-368
[33]吴晓亮,辛萍萍,张志娥,陈晓玲,陶澜,卢新雄.水稻种子室温贮藏最适含水量及其热稳定蛋白的研究.中国农业科学,2006,39(11):2214-2219Wu X L,Xin P P,Zhang Z E,Chen X L,Tao L,Lu X X.Studies on optimum moisture content and heat-stable protein of rice seeds stored at room temperature.Scientia Agricultura Sinica,2006,39(11):2214-2219
[34]周静,辛霞,尹广鹍,陈晓玲,张金梅,卢新雄.大豆种子在不同气候区室温贮藏的适宜含水量与寿命关系研究.大豆科学,2014,33(5):687-690Zhou J,Xin X,Yin G K,Chen X L,Zhang J M,Lu X X.Optimal moisture content and longevity of soybean seeds stored at different climatic zones under ambient condition.Soybean Science,2014,33(5):687-690
[35]辛霞,张金梅,尹广鹍,赵婧,陈晓玲,卢新雄.快速无损检测种子生活力的方法.ZL201510847974.8.2018-03-23Xin X,Zhang J M,Yin G K,Zhao J,Chen X L,Lu XX.A fast,non-invasive technique for seed viability testing.ZL201510847974.8.2018-03-23
[36]辛霞,赵婧,陈晓玲,尹广鹍,张金梅,卢新雄.快速无损检测大豆种子生活力的方法.ZL201510849232.9.2018-02-02Xin X,Zhao J,Chen X L,Yin G K,Zhang J M,Lu X X.Afast,non-invasive technique for soybean seed viability testing.ZL201510849232.9.2018-02-02
[37]辛霞,尹广鹍,陈晓玲,张金梅,赵婧,卢新雄.快速无损检测小麦种子生活力的方法.ZL201510849640.4.2018-08-17Xin X,Yin G K,Chen X L,Zhang J M,Zhao J,Lu X X.Afast,non-invasive technique for wheat seed viability testing.ZL201510849640.4.2018-08-17
[38]Xin X,Wan Y L,Wang W J,Yin G K,ES M L,Lu X X.Areal-time,non-invasive,micro-optrode technique for detecting seed viability by using oxygen influx.Scientific Reports,2013,3(10):3507
[39]王述民,卢新雄,李立会.作物种质资源繁殖更新技术规程.北京:中国农业科学技术出版社,2014Wang S M,Lu X X,Li L H.The technical regulation on regeneration of crop germplasm resources.Beijing:China Agricultural Science and Technology Press,2014
[40]Benson E.E.Cryopreservation of phytodiversity:A critical appraisal of theory practice.Critical Reviews in Plant Sciences,2008,27(3):141-219
[41]Engelmann F.Use of biotechnologies for the conservation of plant biodiversity.In Vitro Cellular and Developmental BiologyPlant,2011,47(1):5-16
[42]Panis B,Swennen R,Rhee J,Roux N.Securing plant genetic resources for perpetuity through cryopreservation.Indian Journal of Plant Genetic Resources,2016,29(3):300-302
[43]International Potato Center.Number of accessions in the CIPCryobank.(2018-08-12)www.cipotato.org/genebankcip/process/cryopreservation-2
[44]Jenderek M M,Reed B M.Cryopreserved storage of clonal germplasm in the USDA National Plant Germplasm System.In Vitro Cellular and Developmental Biology-Plant,2017,53(4):299-308
[45]陈辉,陈晓玲,陈龙清,卢新雄.切花百合离体茎尖玻璃化法超低温保存研究.植物遗传资源学报,2007,8(2):170-173Chen H,Chen X L,Chen L Q,Lu X X.Cryopreservation of shoot tips from in vitro plants of cut flower of lily(Lilium L.)by vitrification method.Journal of Plant Genetic Resources,2007,8(2):170-173
[46]赵艳华,吴雅琴,程和禾,李春敏,陈晓玲,卢新雄.李离体茎尖的超低温保存.园艺学报,2008,35(3):423-426Zhao Y H,Wu Y Q,Cheng H M,Li C M,Chen X L,Lu XX.Cryopreservation of shoot tips from prunus.Acta Horticulturae Sinica,2008,35(3):423-426
[47]刘丽芳.甘薯种质遗传稳定性及超低温保存研究.重庆:西南大学,2009Liu L F.Studies on Genetic Stability and Cryopreservation of Sweet Potato Germplasm.Chongqing:Southwest University,2009
[48]白建明,陈晓玲,卢新雄,郭华春,辛霞,张志娥,辛萍萍.马铃薯茎尖小滴玻璃化法超低温保存及其再生植株的遗传稳定性.园艺学报,2010,37(9):1431-1438Bai J M,Chen X L,Lu X X,Guo H C,Xin X,Zhang Z E,Xin P P.Cryopreservation of in vitro shoot tips of potato by droplet vitrification and genetic stability of regenerated plantlets.Acta Horticulturae Sinica,2010,37(9):1431-1438
[49]李俊慧,何平,陈晓玲,卢新雄,辛霞,张志娥,辛萍萍.香蕉离体茎尖超低温保存研究.植物遗传资源学报,2010,11(1):34-39Li J H,He P,Chen X L,Lu X X,Xin X,Zhang Z E,Xin P P.Cryopreservation of in vitro shoot-tips of banana(Musa spp.).Journal of Plant Genetic Resources,2010,11(1):34-39
[50]Chen X L,Li J H,Xin X,Zhang Z E,Xin P P,Lu X X.Cryopreservation of in vitro-grown apical meristems of Lilium by dropletvitrification.South African Journal of Botany,2011,77(2):397-403
[51]Zhang J M,Zhang X N,Lu X X,Xin X,Yin G K,He J J,Xu Y M,Chen X L.Optimization of droplet-vitrification protocol for carnation genotypes and ultra-structural studies on shoot tips during cryopreservation.Acta Physiologiae Plantarum,2014,36(12):3189-3198
[52]Zhang J M,Huang B,Zhang X N,Volk G M,Zhou Y C,Chen X L.Identification of a highly successful cryopreservation method(droplet-vitrification)for petunia.In Vitro Cellular and Developmental Biology-Plant,2015,51(4):445-451
[53]Zhang J M,Huang B,Lu X X,Volk G M,Xin X,Yin G K,He J J,Chen X L.Cryopreservation of in vitro-grown shoot tips of Chinese medicinal plant Atractylodes macrocephala Koidz.using a droplet-vitrification method.CryoLetters,2015,36(3):195-204
[54]Zhang J M,Han L,Lu X X,Volk G M,Xin X,Yin G K,He JJ,Wang L,Chen X L.Cryopreservation of Jerusalem artichoke cultivars using an improved droplet-vitrification method.Plant Cell and Tissue Organ Culture,2017,128(3):577-587
[55]Zhang J M,Lu X X,Xin X,Yin G K,He J J,Huang B,Jiang D,Chen X L.Cryopreservation of Citrus anthers in the National Crop Genebank of China.In Vitro Cellular and Developmental Biology-Plant,2017,51(4):445-451
[56]张金梅,闫文君,李雪,黄斌,张海晶,赵剑波,姜全,陈晓玲.桃花粉低温和超低温保存方法比较研究.植物遗传资源学报,2017,18(4):670-675Zhang J M,Yan W J,Li X,Huang B,Zhang H J,Zhao J B,Jiang Q,Chen X L.Study on low temperature storage and cryopreservation of pollen for peach germplasm resources.Journal of Plant Genetic Resources,2017,18(4):670-675
[57]陈晓玲,张金梅,辛霞,卢新雄.山葵试管苗离体茎尖包埋干燥超低温保存及再生培养方法,ZL201210211725.6.2014-04-02Chen X L,Zhang J M,Xin X,Lu X X.Methods of cryopreservation and regeneration for in vitro-grown shoot tips of wasabi.ZL201210211725.6.2014-04-02
[58]陈晓玲,张金梅,卢新雄,辛霞,尹广鹍,何娟娟.桑树休眠芽段的超低温保存及嫁接复活方法,ZL201410366753.4.2016-06-11Chen X L,Zhang J M,Lu X X,Xin X,Yin G K,He J J.Methods of cryopreservation and grafting regeneration for mulberry dormant bud.ZL201410366753.4.2016-06-11
[59]Huang B,Zhang J M,Chen X L,Xin X,Yin G K,He J J,Lu X X,Zhou Y C.Oxidative damage and antioxidative indicators in 48 h germinated rice embryos during the vitrificationcryopreservation procedure.Plant Cell Reports,2018,37:1325-1342
[60]李运合,李玉生,张金梅,陈晓玲,赵艳华,吴永杰.芒果细胞超低温保存存活机理.植物生理学报,2016,52(10):1474-1480Li Y H,Li Y S,Zhang J M,Chen X L,Zhao Y H,Wu Y J.The survival mechanism of mango cells during cryopreservation.Plant Physiology Journal,2016,52(10):1474-1480
[61]张海晶,张金梅,辛霞,尹广鹍,何娟娟,卢新雄,周元昌,陈晓玲.马铃薯茎尖超低温保存流程TTC活力响应.植物遗传资源学报,2015,16(3):555-560Zhang H J,Zhang J M,Xin X,Yin G K,He J J,Lu X X,Zhou Y C,Chen X L.Study on potato shoot tips vitality responses during cryopreservation by TTC staining method.Journal of Plant Genetic Resources,2015,16(3):555-560
[62]Zhang J M,Xin X,Yin G K,Lu X X,Chen X L.In vitro conservation and cryopreservation in National Genebank of China.Acta Horticulturae,2014,1039(1039):309-317
[63]陈晓玲,张金梅,辛霞,黄斌,卢新雄.植物种质资源超低温保存现状及其研究进展.植物遗传资源学报,2013,14(3):414-427Chen X L,Zhang J M,Xin X,Huang B,Lu X X.Progress on cryopreservation state and research of plant germplasm resources.Journal of Plant Genetic Resources,2013,14(3):414-427
[64]何娟娟,卢新雄,栗钊,辛霞.8种粮食和蔬菜作物种子短期节能储藏研究.中国种业,2016(9):57-60He J J,Lu X X,Li Z,Xin X.Short-term storage of 8 crop and vegetable seeds with an energy-saving method.China Seed Industry,2016(9):57-60
[65]卢新雄,陈叔平,刘旭.农作物种质资源保存技术规程.北京:中国农业出版社,2008Lu X X,Chen S P,Liu X.The technical regulation on conservation of crop germplasm resources,Beijing:China Agriculture Press,2008
[66]卢新雄.农业种质库的设计与建设要求探讨.农业工程学报,2003,19(6):252-255Lu X X.Design and construction of agricultural germplasm genebank.Transactions of The Chinese Society of Agricultural Engineering,2003,19(6):252-255
[67]郭颖荷,卢新雄,彭高军.中国低温种质库建造技术调查分析.农业工程学报,2005,21(4):186-190Guo Y H,Lu X X,Peng G J.Investigation and analysis of the building technology of low temperature germplasm genebanks of China.Transactions of The Chinese Society of Agricultural Engineering,2005,21(4):186-190
[68]中华人民共和国农业农村部,中华人民共和国国家发展和改革委员会,中华人民共和国科学技术部.全国农作物种质资源保护与利用中长期发展规划(2015-2030年).(2015-04-29)[2018-08-15]http://www.moa.gov.cn/nybgb/2015/si/201711/t20171129_6134098.htm Ministry of Agriculture of the People's Republic of China,National Development and Reform Commission of the People's Republic of China,People's Republic of China Ministry of Science and Technology.National Medium-and Long-term Development Plan for the Protection and Utilization of Crop Germplasm Resources(2015-2030).(2015-04-29)[2018-08-15]http://www.moa.gov.cn/nybgb/2015/si/201711/t20171129_6134098.htm