大花萱草EMS离体诱变和抗叶枯病突变体的筛选
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摘要
利用甲基磺酸乙酯(EMS)诱变处理大花萱草愈伤组织和分化芽,经过萱草叶枯病菌毒素离体筛选获得抗病愈伤组织和分化芽突变体。以不同浓度剂量EMS诱变处理大花萱草子房诱导的愈伤组织和分化芽,用经过毒力检测的萱草叶枯病菌毒素粗提液进行离体筛选,通过在继代培养基中添加毒素粗提液,对愈伤组织进行浓度递增法和分化芽一步筛选法,获得抗叶枯病突变体材料。结果表明,0.50%~0.75%EMS处理愈伤组织60 min,0.75%~1.00%EMS处理分化芽60 min,分别获得半致死剂量效应。对存活的愈伤组织进行病菌毒素浓度递增逐步筛选(10%30天~20%30天),分化芽用40%的毒素液一步法筛选21天。经定向筛选,初步获得抗叶枯病分化芽突变体103株,有待进一步进行抗病鉴定。愈伤组织未能获得抗病突变体再生植株。
The calli and differentiation buds from ovary of Hemerocallis hybrida were mutagenized in vitro bydifferent concentrations of EMS(ethyl methane sulfonate) and screened by toxin extracts after the toxicitytesting of Kabatiella microsticta on Hemerocallis hybrida. The results showed that 0.50%-0.75% EMS treatedcallus 60 min, 0.75%-1.00% EMS treated differentiation bud 60 min, and obtained semi lethal dose effectrespectively. The surviving calli were gradually screened for bacterial toxin concentration(10%, 30 days-20%, 30 days), and the surviving differentiated buds were screened for 21 days with 40% toxin, one stepsolution. 103 mutants of leaf blight from bud differentiation were obtained by directional screening, and furtheridentification was needed. Calli failed to obtain resistant mutant plants.
The calli and differentiation buds from ovary of Hemerocallis hybrida were mutagenized in vitro bydifferent concentrations of EMS(ethyl methane sulfonate) and screened by toxin extracts after the toxicitytesting of Kabatiella microsticta on Hemerocallis hybrida. The results showed that 0.50%-0.75% EMS treatedcallus 60 min, 0.75%-1.00% EMS treated differentiation bud 60 min, and obtained semi lethal dose effectrespectively. The surviving calli were gradually screened for bacterial toxin concentration(10%, 30 days-20%, 30 days), and the surviving differentiated buds were screened for 21 days with 40% toxin, one stepsolution. 103 mutants of leaf blight from bud differentiation were obtained by directional screening, and furtheridentification was needed. Calli failed to obtain resistant mutant plants.
引文
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[26]林丛发.EMS诱导太子参抗性突变体及分子检测[J].核农学通报,2014,28(7):1155-1161.
[27]徐美隆,张怀渝,唐宗祥,等.EMS对丽格海棠离体诱变的生物学效应[J].核农学报,2007,21(6):577-596.
[28]刘君绍,田时炳,皮伟,等.茄子抗黄萎病突变体离体筛选[J].西南农业学报,2003,16(4):102-106.
[2]白庆荣,韩双,赵莹,等.萱草叶枯病菌生物学特性及对药剂敏感性研究[J].园艺学报,2013,40(12):2513-2519.
[3]陈丽飞,董然.萱草属植物研究进展[J].北方园艺,2007(6):66-69.
[4]张伟丽,刘凤民,许旭丹.组培法筛选高抗盐大花萱草品种的初步研究[J].北方园艺,2009(11):184-186.
[5]金立敏,周玉珍,郭志海,等.大花萱草自交、杂交及不同栽培环境结实率比较[J].湖北农业科学,2012,51(13):2760-2763.
[6]雷媛.不同倍性大花萱草杂交亲和性的研究[D].太谷:山西农业大学,2013:16-21.
[7]郑楠,王霞霞,曹冬梅.农杆菌介导萱草植株CHS基因的转化[J].中国园艺文摘,2015,2:5-11.
[8]蔡海燕,温立柱,郑成淑,等.EMS诱发菊花突变类型及重要性状的分子鉴定[J].山东农业大学学报:自然科学版,2013,44(2):171-175.
[9]罗静,周厚成,王永清,等.EMS离体诱变及抗草莓灰霉病愈伤组织的筛选[J].核农学报,2009,23(1):90-94.
[10]杨媚,舒灿伟,陈建仪,等.利用甲基磺酸乙酯诱变筛选香蕉抗毒素突变体[J].园艺学报,2012,39(8):1465-1470.
[11]杨广东,郭勤平,马树斌,等.大白菜抗黑斑病突变株筛选技术研究[J].中国农学通报,2000,16(6):57-58.
[12]殷冬梅,杨秋云,杨海棠,等.花生突变体的EMS诱变及分子检测[J].中国农学通报,2009,25(05):53-56.
[13]Joong Ho Lee.Selection of stable mutants from cultured rice anthers treated with ethylmethane sulfonic acid[J].Plant Cell Tissue and Culture,2002,71(2):165-171.
[14]安学丽,蔡一林,王久光,等.甲基磺酸乙酯(EMS)对玉米自交系诱变效应的研究[J].玉米科学,2003,11(3):74-84.
[15]杨丽莉,王德平,张晓,等.以子房为外植体的金娃娃萱草组织培养技术的研究[J].中国农学通报,2012,28(25):184-190.
[16]陈超,王桂兰,乔永旭,等.蝴蝶兰类圆球茎的化学诱变试验[J].核农学报,2006,20(2):99-102.
[17]吴金平,顾玉成,万进,等.魔芋抗软腐病突变体筛选的初步研究[J].华中农业大学学报,2005,24(5):448-450.
[18]Lyons R E.About Daylilies[J].The Virgiain Gardener Newsletter,1993,12:1.
[19]Rhoads A F.Leaf-streak disease of daylily observed in Pennsylvania[J].Plant Disease Reporter,1974,58:102.
[20]Stephanie Porter,Zu Dienle Tan.Daylily Leaf Strenk(Aureobasidium microstictum)[J].Home,Yard&Garden Pest Newsletter,2012,8.
[21]William W Kirk,Rob Schafer,Devan Berry.Control of Leaf streak on daylilies with fungicides[J].Plant Diseases and Disease Management,2002:23-24.
[22]Dicklow M B.Daylily streak and daylily rust[J].Floral Notes Newsletter,2011,23(6):5.
[23]金立敏,张文婧,霍尧,等.萱草不同品种杂交亲和性及其种子萌芽力研究[J].安徽农业科学,2009,37(6):2471-2472.
[24]徐冠仁.植物诱变育种学[M].北京:中国农业出版社,1996:1-2.
[25]朴铁夫,原亚萍,郭筑英,等.EMS对水稻成熟胚愈伤组织生长和植株分化率的影响[J].核农学报,1995,16(2):56-58.
[26]林丛发.EMS诱导太子参抗性突变体及分子检测[J].核农学通报,2014,28(7):1155-1161.
[27]徐美隆,张怀渝,唐宗祥,等.EMS对丽格海棠离体诱变的生物学效应[J].核农学报,2007,21(6):577-596.
[28]刘君绍,田时炳,皮伟,等.茄子抗黄萎病突变体离体筛选[J].西南农业学报,2003,16(4):102-106.