“寒富”苹果败育胚珠挽救影响因子研究
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摘要
以"寒富"苹果子房及其败育胚珠为试材,研究不同接种时间、基本培养基种类、不同激素质量浓度和氨基酸组份、培养基相态组成、光照和暗培养对子房离体培养及败育胚珠发育率的影响。结果表明,在大蕾期和子房直径0.3cm(开花后5d)时最适宜胚珠生长的培养基为Nitsch培养基+叶酸5mg/L+IAA 0.5mg/L+6-BA 1mg/L+GA32mg/L,而在开花后10d,适宜败育胚珠生长的培养基为Nitsch培养基+叶酸5mg/L+IAA 1mg/L+6-BA 1mg/L+GA32mg/L,其挽救率最高,达到80%;在"寒富"苹果胚珠败育后期,起主要作用的氨基酸有半胱氨酸和谷氨酸,多种氨基酸培养效果高于任何一种单一氨基酸培养。固液双相培养基对于败育胚珠的培养效果明显优于单独相态培养;败育胚珠在连续黑暗条件下培养其发育率高于光暗交替培养,并且经TTC检测发现暗培养下胚珠的生活力显著高于光暗交替。
In this experiment,ovaries and abortive ovules of "Hanfu" apple were used to investigate the effect of different inoculation time,different basic media,different hormone concentrations and amino acid species,different medium phases,and light/dark condition on development rate of abortive ovules and ovaries culture in vitro.The results indicated that the most suitable medium for ovules growth was Nitsch + folic acid 5 mg/L + IAA 0.5 mg/L + 6-BA 1 mg/L + GA3 2 mg/L at big bud stage and ovary diameter was 0.3 cm(5 days after flower),while the most suitable medium for ovules growth was Nitsch + folic acid 5 mg/L + IAA 1 mg/L + 6-BA 1 mg/L + GA3 2 mg/L at 10 days after flower.The rescue rate with this formula was the highest,reached to 80%.Moreover,in the late stage of abortive ovules development,cysteine and glutamate played a major role in ovules rescue,the training effect of various amino acids was better than single amino acid.Solid-liquid double phase medium had the most positive effect on abortive ovules development.The development rate of abortive ovules under dark condition was higher than under light and dark alternative culture condition.The viability of ovules,which were cultured under dark condition and detected by TTC,was significantly higher than that under light and dark alternative culture condition.
In this experiment,ovaries and abortive ovules of "Hanfu" apple were used to investigate the effect of different inoculation time,different basic media,different hormone concentrations and amino acid species,different medium phases,and light/dark condition on development rate of abortive ovules and ovaries culture in vitro.The results indicated that the most suitable medium for ovules growth was Nitsch + folic acid 5 mg/L + IAA 0.5 mg/L + 6-BA 1 mg/L + GA3 2 mg/L at big bud stage and ovary diameter was 0.3 cm(5 days after flower),while the most suitable medium for ovules growth was Nitsch + folic acid 5 mg/L + IAA 1 mg/L + 6-BA 1 mg/L + GA3 2 mg/L at 10 days after flower.The rescue rate with this formula was the highest,reached to 80%.Moreover,in the late stage of abortive ovules development,cysteine and glutamate played a major role in ovules rescue,the training effect of various amino acids was better than single amino acid.Solid-liquid double phase medium had the most positive effect on abortive ovules development.The development rate of abortive ovules under dark condition was higher than under light and dark alternative culture condition.The viability of ovules,which were cultured under dark condition and detected by TTC,was significantly higher than that under light and dark alternative culture condition.
引文
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[5]ZHAO Deying(赵德英),LIU Guocheng(刘国成),LDe-guo(吕德国),et al.Evaluation of the specific characters ofHanfu apple cultivar[J].Journal of Fruit Science(果树学报),2009,26(1):6-12(in Chinese with English abstract).
[6]KONG Xiangsheng(孔祥生),YI Xianfeng(易现峰).TheExperimental Technique of Phytophysiology(植物生理学实验技术)[M].Beijing:China Agriculture Press,2008,236(in Chinese).
[7]XIA Peibei(夏培蓓),WU Xinyu(伍新宇),WU Yuxia(吴玉霞),et al.A study on technique for vestigal ovules cultureof white munake grape[J].Xinjiang Agricultural Sciences(新疆农业科学),2011,48(7):1201-1204(in Chinese withEnglish abstract).
[8]Ponce M T,Agüero C B,Gregori M T,et al.Factors affect-ing the development of stenospermic grape(Vitis viniferaL.)embryos cultured in vitro[J].Acta Horticulturae,2000,528(2):667-671.
[9]NIU Ruxuan(牛茹萱),ZHANG Jianxia(张剑侠),WANGYuejin(王跃进),et al.Research on the embryo rescue ofstenospemocarpic seedless grape[J].Northern Horticul-ture(北方园艺),2012(7):1-4(in Chinese with Englishabstract).
[10]ZHOU Qingyuan(周清元),LI Jiana(李加纳),YIN Jiam-ing(殷家明),et al.Preliminary studies on interspecifichybridization between Brassica campestris and B.oleraceavar.aceaphalaⅠ.The effect of the time of overay takingon seed forming in in vitro culture[J].Journal of South-west Agricultural University:Natural Science Edition(西南农业大学学报:自然科学版),2003,25(6):495-509(inChinese with English abstract).
[11]Spiegle-Roy P,Sahar N,Baron J,et al.In vitro culture andplant formation from grape cultivars with abortic ovulesand seeds[J].Journal of the American Society for Horti-cultural Sciences,1985,110(1):109-112.
[12]Tian L L,Wang Y J,Niu L,et al.Breeding of disease-re-sistant seedless grapes using Chinese wild Vitis spp.I.Invitro embryo rescue and plant development[J].ScientiaHorticulturae,2008,117(2):136-141.
[13]Nalini M.Ovule and embryo culture to obtain hybrids frominterspecific incompatible pollinations in chickpea[J].Eu-phytica,1999,110(1):1-6.
[14]XIA Qizhong(夏启中),ZHANG Xianlong(张献龙),NIEYichun(聂以春),et al.Withdrawal of exogenous auxininduces programmed cell death o f cotton embryogenic sus-pension cultures[J].Journal of Plant Physiology andMdecular Biology(植物生理与分子生物学学报),2005,31(1):78-84(in Chinese with English abstract).
[15]LIU Sanjun(刘三军),KUAI Chuanhua(蒯传化),YU Qia-oli(于巧丽),et al.Research and application of the inherit-ance of seedlessness and embryo rescue techniques in seed-less grape breeding[J].Journal of Fruit Science(果树学报),2009,26(1):71-76(in Chinese with English ab-stract).
[16]Emershad R L,Ramming D W.In ovulo embryo culture ofVitis vinifera L.cv.“Thompson Seedless”[J].AmericanJournal of Botany,1984,71(6):873-877.
[17]Fuller R J,Carman J G,Hess J R.Nutrient and hormonelevels in cotton ovules during embryony[J].Plant Cell TissOrgan Cult,2009,99(2):183-192.
[18]Burger P,Trautmann I A.Manipulations of ovules to im-prove in vitro development of Vitis vinifera L.embryos[J].Acta Horticulturae,2000,528:613-619.
[19]SUN Liang(孙亮),LENG Ping(冷平).Apple-pearhybrid and in vitro rescue culture of zygotic embryo[J].Journal of China Agricultural University(中国农业大学学报),2008,13(2):25-29(in Chinese with English ab-stract).
[20]Uma S,Lakshmi S,Saraswathi M S,et al.Embryo rescueand plant regeneration in banana(Musa spp.)[J].PlantCell Tiss Organ Cult,2011,105(1):105-111.
[21]Esin A,Hatice I,Munevver G,et al.Comparative evalua-tion of different embryo rescue techniques on parthenoge-netic melon(Cucumis melo L.)fruits induced with irradi-ated pollen[J].African Journal of Biotechnology,2010,9(33):5347-5356.
[22]Bhuyar S A,Sakhare S B,Patil B R,et al.Embryo rescuestudies in interspecific hybrids of cotton[J].Annals ofPlant Physiology,2009,23(2):272-273.
[2]TANG Dongmei(唐冬梅),WANG Yuejin(王跃进),ZHAORonghua(赵荣华),et al.Factors influencing embryo de-velopment in embryo rescue of seedless grapes[J].Sci Ag-ric Sin(中国农业科学),2009,42(7):2449-2457(in Chi-nese with English abstract).
[3]GUO Xiuwu(郭修武),GUO Yinshan(郭印山),ZHANGHaie(张海娥),et al.Effects of culture medium and inocu-lating date on embryo rescue of seedless grape[J].ActaHorticulturae Sinica(园艺学报),2007,34(2):329-332(inChinese with English abstract).
[4]SHU Huairui(束怀瑞).Apple Science(苹果学)[M].Bei-jing:China Agriculture Press,1999,310(in Chinese).
[5]ZHAO Deying(赵德英),LIU Guocheng(刘国成),LDe-guo(吕德国),et al.Evaluation of the specific characters ofHanfu apple cultivar[J].Journal of Fruit Science(果树学报),2009,26(1):6-12(in Chinese with English abstract).
[6]KONG Xiangsheng(孔祥生),YI Xianfeng(易现峰).TheExperimental Technique of Phytophysiology(植物生理学实验技术)[M].Beijing:China Agriculture Press,2008,236(in Chinese).
[7]XIA Peibei(夏培蓓),WU Xinyu(伍新宇),WU Yuxia(吴玉霞),et al.A study on technique for vestigal ovules cultureof white munake grape[J].Xinjiang Agricultural Sciences(新疆农业科学),2011,48(7):1201-1204(in Chinese withEnglish abstract).
[8]Ponce M T,Agüero C B,Gregori M T,et al.Factors affect-ing the development of stenospermic grape(Vitis viniferaL.)embryos cultured in vitro[J].Acta Horticulturae,2000,528(2):667-671.
[9]NIU Ruxuan(牛茹萱),ZHANG Jianxia(张剑侠),WANGYuejin(王跃进),et al.Research on the embryo rescue ofstenospemocarpic seedless grape[J].Northern Horticul-ture(北方园艺),2012(7):1-4(in Chinese with Englishabstract).
[10]ZHOU Qingyuan(周清元),LI Jiana(李加纳),YIN Jiam-ing(殷家明),et al.Preliminary studies on interspecifichybridization between Brassica campestris and B.oleraceavar.aceaphalaⅠ.The effect of the time of overay takingon seed forming in in vitro culture[J].Journal of South-west Agricultural University:Natural Science Edition(西南农业大学学报:自然科学版),2003,25(6):495-509(inChinese with English abstract).
[11]Spiegle-Roy P,Sahar N,Baron J,et al.In vitro culture andplant formation from grape cultivars with abortic ovulesand seeds[J].Journal of the American Society for Horti-cultural Sciences,1985,110(1):109-112.
[12]Tian L L,Wang Y J,Niu L,et al.Breeding of disease-re-sistant seedless grapes using Chinese wild Vitis spp.I.Invitro embryo rescue and plant development[J].ScientiaHorticulturae,2008,117(2):136-141.
[13]Nalini M.Ovule and embryo culture to obtain hybrids frominterspecific incompatible pollinations in chickpea[J].Eu-phytica,1999,110(1):1-6.
[14]XIA Qizhong(夏启中),ZHANG Xianlong(张献龙),NIEYichun(聂以春),et al.Withdrawal of exogenous auxininduces programmed cell death o f cotton embryogenic sus-pension cultures[J].Journal of Plant Physiology andMdecular Biology(植物生理与分子生物学学报),2005,31(1):78-84(in Chinese with English abstract).
[15]LIU Sanjun(刘三军),KUAI Chuanhua(蒯传化),YU Qia-oli(于巧丽),et al.Research and application of the inherit-ance of seedlessness and embryo rescue techniques in seed-less grape breeding[J].Journal of Fruit Science(果树学报),2009,26(1):71-76(in Chinese with English ab-stract).
[16]Emershad R L,Ramming D W.In ovulo embryo culture ofVitis vinifera L.cv.“Thompson Seedless”[J].AmericanJournal of Botany,1984,71(6):873-877.
[17]Fuller R J,Carman J G,Hess J R.Nutrient and hormonelevels in cotton ovules during embryony[J].Plant Cell TissOrgan Cult,2009,99(2):183-192.
[18]Burger P,Trautmann I A.Manipulations of ovules to im-prove in vitro development of Vitis vinifera L.embryos[J].Acta Horticulturae,2000,528:613-619.
[19]SUN Liang(孙亮),LENG Ping(冷平).Apple-pearhybrid and in vitro rescue culture of zygotic embryo[J].Journal of China Agricultural University(中国农业大学学报),2008,13(2):25-29(in Chinese with English ab-stract).
[20]Uma S,Lakshmi S,Saraswathi M S,et al.Embryo rescueand plant regeneration in banana(Musa spp.)[J].PlantCell Tiss Organ Cult,2011,105(1):105-111.
[21]Esin A,Hatice I,Munevver G,et al.Comparative evalua-tion of different embryo rescue techniques on parthenoge-netic melon(Cucumis melo L.)fruits induced with irradi-ated pollen[J].African Journal of Biotechnology,2010,9(33):5347-5356.
[22]Bhuyar S A,Sakhare S B,Patil B R,et al.Embryo rescuestudies in interspecific hybrids of cotton[J].Annals ofPlant Physiology,2009,23(2):272-273.