杏胚珠培养及幼胚子叶不定芽诱导研究Ovule culture and adventitious bud regeneration from immature cotyledons of apricot
楚宗丽;王力荣;朱更瑞;冯义彬;方伟超;
摘要(Abstract):
以盛花后25、35、50、55、60d的凯特杏胚珠为外植体,研究胚珠培养的适宜条件;选择发育一致的幼胚子叶,研究不定芽的发生条件。结果表明,SH培养基是凯特杏胚珠培养的适宜培养基;盛花后25d的凯特杏胚珠(PF=0),仅采用生长培养不足以发育成可见胚,试验采用诱导方式获得了可见胚,诱导出胚率为80%;以MS为基本培养基,BA与NAA结合诱导能力强;但这种胚小且瘦弱,继续生长培养,利用幼胚子叶诱导不定芽。在幼胚子叶诱导不定芽方面,盛花后25、35d取样的凯特杏胚珠,培养后获得幼胚子叶,诱导不定芽的适宜TDZ质量浓度为1.25mg/L,诱导出芽率分别为85.00%、80.00%;盛花后50、55d取样的材料,适宜TDZ质量浓度为2.50mg/L,出芽率分别为65.00%、60.00%,而盛花后60d取样的材料,适宜TDZ质量浓度为5.0mg/L,出芽率为61.11%,不定芽主要发生在子叶的近轴面、胚轴连接处及子叶近轴端两侧;在相同培养基中,25℃进行21d暗培养,盛花后35、50、55、60d材料出芽率分别为72.22%、65.00%、60.00%、55.56%;4℃下28d暗培养,35、50、55、60d依次为59.09%,71.42%、77.78%、86.36%;对不定芽进行伸长、生根与快繁培养,获得了健康正常的植株。通过以上研究,建立了凯特杏幼胚4级培养体系,即(1)胚珠培养,获得可见胚(1级培养);(2)幼胚生长培养(胚培养),获得了健壮幼胚(2级培养);(3)幼胚子叶再生,获得不定芽(3级培养);(4)不定芽组培苗快繁(4级培养)。
关键词(KeyWords): 杏;胚珠培养;幼胚子叶;不定芽诱导
基金项目(Foundation):
作者(Author): 楚宗丽;王力荣;朱更瑞;冯义彬;方伟超;
Email:
DOI:
参考文献(References):
- [1]YANG H H,CHEN X S,FENG B C,LIU H F,ZHENG Z.Creating new germplasm by distant hybridization in stone fruitsⅡ.Embryo rescue and hybrid identification between plum and apricot[J].Sci-ence Agricultura Sinica,2004,37(8):1203-1207.(in Chinese)杨红花,陈学森,冯宝春,刘焕芳,郑洲.利用远缘杂交创造核果类果树新种质的研究II.李、杏远缘杂种胚挽救及杂种鉴定[J].中国农业科学,2004,37(8):1203-1207.
- [2]RAMMING D W.The use of embryo culture in breeding[J].Hortscience,1990,25(4):393-398.
- [3]HAMMERSCHLAG F A,BAUCHAN G,SCORZA R.Regeneration of peach from callus derived from immature embryos[J].Theor Appl Genet,1985,70:248-251.
- [4]BHANSALI R R,DRIVER J A,DURZAN D J.Somatic embryogene-sis in cell suspension cultures ofPrunus persica(L)[J].Joural of Hor-ticultural Science,1991,66(5):601-605.
- [5]LIU Y S,LI X J.Embryo culture and application of peach[J].North-ern Fruits ofChina,1993(2):3-5.(in Chinese)刘用生,李秀菊.桃胚胎培养及其应用[J].北方果树,1993(2):3-5.
- [6]ZHUANG E J,YAO Q,WU Y L,WANG D C.Studies on in-ovule embyro culture techniques of early–mature and very early-mature peach[J].Acta Horticulture Sinica,1991,18(4):303-308.(in Chi-nese)庄恩及,姚强,吴钰良,王得春.早熟和特早熟桃胚珠培养研究[J].园艺学报,1991,18(4):303-308.
- [7]WANG Y Z,ZHANG D P,YANG L,YAN A L,SHI H.Study on the techniques of in vitro embryo culture with immature embryo of Apri-cot[J].Journal ofAgricultural Biotechnology,2000,8(4):389-391.(in Chinese)王玉柱,张大鹏,杨丽,阎爱玲,石宏.未成熟杏离体胚培养技术的研究[J].农业生物技术学报,2000,8(4):389-391.
- [8]YAN G H,ZHOU Y.Plant regeneration from excised immature em-bryos of peach(Prunus persica L)[J].Acta Horticulture Sinica,2002,29(5):480-482.(in Chinese)阎国华,周宇.桃幼胚离体培养再生植株的研究[J].园艺学报,2002,29(5):480-482.
- [9]WU Y J,ZHANG S L,ZHANG L L,SHEN G Q,WU D J.Shoots re-generation from immature cotyledons of peach(Prunus persica L.Batsch)[J].Journal of Zhejiang University,2003,29(1):93-96.(in Chinese)吴延军,张上隆,张岚岚,沈淦卿,吴大军.桃幼胚子叶不定芽发生的初步研究[J].浙江大学学报,2003,29(1):93-96.
- [10]SUN Y F,HAN MY,FAN C H,ZHANG MR,WANG S L.Influen-tial factors for nectarine cotyledon regeneration[J].Journal of North-west Sci Tech Univ of Agri and For:Nat Sci Ed,2004,32(4):66-70.(in Chinese)孙跃峰,韩明玉,范崇辉,张满让,王淑莉.油桃子叶再生植株影响因子研究[J].西北农林科技大学学报:自然科学版,2004,32(4):66-70.
- [11]ZHU J J,WU H M,WANG Z H.Studies on the technique of embryo culture of early-mature peachs[J].Journal of Jangsu Agri Sci,1983(9):33-35.(in Chinese)朱际君,吴鹤鸣,汪祖华.桃早熟品种种胚培养技术研究[J].江苏农业科学,1983(9):33-35.
- [12]CHAPORRO J X.Culture date and germination procedure affects succes of neclafine ovule and embryo culture[J].Fruit Varielies Journal,1994,48(3):173-175.
- [13]RAMMING D W.In ovule embryo of early-maturing Prunus[J].Hortscience,1985,20(3):419-420.
- [14]MANTE S,SCORZA R,CORDTS J M.Plant regeneration from cotyledons of Prunus persica,Prunus domestica,and Prunus cera-sus[J].Plant Cell,Tissue and Organ Culture,1989,19:1-11.
- [15]SUN Q R,SUN H Y,SHI Y P,LIU Q Z.Shoot induction from juve-nile cotyledons of early apricot[J].Plant Physiology Communica-tions,2001,37(1):38-39.(in Chinese)孙清荣,孙洪雁,石荫坪,刘庆忠.早熟杏幼胚子叶不定芽诱导[J].植物生理学通讯,2001,37(1):38-39.
- [16]HAN MY,ZHANG MR,TIAN Z S,SUN Y F,TIAN Y M,WANG A Z,ZHAO C P.Plant regeneration with in vitro culture from cotyle-dons and hypocotyls of earlyripen nectarines(Prunus persicav nec-tarina)[J].Acta Horticulture Sinica,2006,33(5):957-962.(in Chi-nese)韩明玉,张满让,田增胜,孙跃峰,田玉命,王安柱,赵彩平.早熟油桃子叶和胚轴离体培养再生植株的研究[J].园艺学报,2006,33(5):957-962.
- [17]WANGYM,CAOZY,LI W,LI S,ZHOUWT,LI WB.Studyon the induction and plantlet regerenation from ovule of grape[J].Journal of Gansu Agricultural University,2005,40(2):167-172.(in Chinese)王雅梅,曹孜义,李唯,李胜,周万涛,李文彬.葡萄胚珠诱导成苗的研究[J].甘肃农业大学学报,2005,40(2):167-172.
- [18]WU Y J,ZHANG S L,XIE M,CHEN J W,QIN Y H,QIN Q P.Plantlets regeneration from immature embryos and cotyledons of peach[J].Scientia Silvae Sinicae,2005,41(5):45-50.(in Chinese)吴延军,张上隆,谢鸣,陈俊伟,秦永华,秦巧平.桃幼胚及幼胚子叶再生的研究[J].林业科学,2005,41(5):45-50.