摘要
【目的】探究胚挽救无核葡萄畸形苗发生原因及转变成正常葡萄试管苗的方法,提高无核葡萄胚挽救育种效率。【方法】‘无核白’(‘Thompson seedless grape’)、‘火焰无核’(‘Flame seedless grape’)、‘赫什无核’(‘Heshi seedless grape’)及‘红宝石无核’(‘Ruby seedless grape’)自交以及和不同父本杂交后进行胚挽救,研究不同亲本基因型和不同胚萌发培养基对胚挽救畸形苗产生的影响。通过不同培养基及培养方式离体培养胚挽救畸形苗,进一步研究胚挽救畸形苗转变成正常葡萄试管苗的方法。【结果】‘无核白’和‘赫什无核’作母本,用MS培养基培养易产生胚挽救畸形苗。畸形胚萌发苗可总结为5大类:白化苗、玻璃化苗、徒长苗、子叶扭曲无根苗和无生长点的有根苗。转接在WPM+6-BA 2.0 mg·L^-1+IAA 2.0 mg·L^-1+琼脂6 g·L^-1+蔗糖20 g·L^-1+活性炭2.0 g·L^-1培养基上培养,每隔14 d进行继代转接,将畸形苗转变成正常葡萄试管苗效果较好。【结论】无核葡萄胚挽救产生的畸形苗及时转接到不同培养基上或者采取合适方式进行继代培养,可以将胚挽救畸形苗转变成正常葡萄试管苗。
【Objective】The objective of this study was to explore the causes of abnormal plantlets in the process of embryo rescue of seedless grapes and to find effective measures to prevent the occurrence abnormal seedlings.With embryo rescue,efficiency of seedless grape breeding can be improved.【Methods】Abnormal seedlings were used as the test materials.These abnormal seedlings developed from embryos rescued in vitro from self-pollination of four seedless grape(Vitis vinifera L.)cultivars‘(Thompson seedless’‘Flame seedless’‘Heshi seedless’and‘Ruby seedless’)and embryos from different crosses of seedless grapes.The effects of different parent genotypes and different embryo germination media on the emergence of abnormal seedlings rescued from seedless grape embryos were studied.The 4 different parent genotypes were‘Thompson seedless’‘Flame seedless’‘Heshi seedless’and‘Ruby seedless’.The 5 different media were MS,MM3,MM4,ER,and modified MM3.Different methods for transforming abnormal seedlings into normal seedlings were studied in vitro.The methods included 4 different culture media and different subculture durations.The media included WPM(W1),WPM+6-BA 2.0mg·L^-1(W2),WPM+6-BA 2.0 mg·L^-1+IAA 2.0 mg·L^-1(W3)and WPM+IAA 2.0 mg·L^-1(W4).These media were all added with agar 6 g·L^-1+sucrose 20 g·L^-1+activated carbon 2.0 g·L^-1.Normal seedlings were observed and their number was counted after 45 days.The different subculture durations were 7 d,14 d,21 d and 28 d,and normal seedlings were counted after 60 days.【Results】It was difficult to rescue‘Thompson seedless’embryos.The proportion of abnormal seedlings from rescued embryos of‘Thompson seedless’and‘Heshi seedless’was higher that from the other cultivars.The 5 embryos development media were used to culture in vitro embryos from self-pollination of the four seedless grape cultivars.The percentage of abnormal seedlings was always highest on the MS medium.The percentage of abnormal seedlings of‘Thompson seedless’was 33.3%,‘Flame seedless’37.5%,‘Heshi seedless’33.3%,and‘Ruby seedless’23.1%.The abnormal germination embryo rate of the 5 media followed a pattern of modified MM3<MM3<ER<MM4<MS.Therefore,MS medium was not suitable to be used as the rescuing medium for seedless grape embryos.The abnormal grape seedlings were classified into 5 types:(1)albino seedlings,(2)vitrified seedlings,(3)seedlings with excessive growth,(4)plantlets with contorted cotyledon and no roots,and(5)plantlet without shoot tip.All these different types of abnormal seedlings could be transformed into normal seedlings using appropriate media.The percent of seedlings transformed to normal ones was highest(24.6%)when cultured on WPM+6-BA 2.0 mg·L^-1+IAA 2.0 mg·L^-1,and lowest(8.1%)when cultured on WPM.After leggy seedlings were cut for subculture every 14 days,the percentage of normal seedlings was higher(33.3%)than other intervals.Albino and vitrified seedlings were not able to turn into normal seedlings.Finally,these normal seedlings were hardened and transplanted in the field,and the operation steps for seedling hardening and transplantation into fields were summarized.【Conclusion】Abnormal plantlet occurrence from rescued embryos of‘Thompson seedless’,‘Heshi seedless’was high,especially cultured in MS medium.WPM+6-BA 2.0 mg·L^-1+IAA 2.0 mg·L^-1+agar 6 g·L^-1+sucrose 20 g·L^-1+activated carbon 2.0 g·L^-1 was suitable to normalize the abnormal seedlings.For leggy plantlets,the transformation rate was high with a subculture duration of 14 days,reaching 33.3%.The abnormal plantlets produced by embryo rescue of seedless grapes can be transferred to normal plantlets on proper media and subcultured in a proper way.The abnormal plantlets except albino plantlets and vitrified plantlets could be transformed into normal plantlets.
作者
李桂荣
全冉
程珊珊
侯小进
蔡祖国
扈惠灵
LI Guirong;QUAN Ran;CHENG Shanshan;HOU Xiaojin;CAI Zuguo;HU Huiling(School of Horticulture and Landscape Architecture,Henan Institute of Science and Technology,Xinxiang 453003,Henan,China;Henan Province Engineering Research Center of Horticultural Plant Resource Utilization and Germplasm Enhancement,Xinxiang 453003,Henan,China)
出处
《果树学报》
CAS
CSCD
北大核心
2020年第6期819-829,共11页
Journal of Fruit Science
基金
河南省自然科学基金(182300410045)。
关键词
无核葡萄
胚挽救
畸形胚挽救苗
正常葡萄试管苗
驯化移栽
Seedless grape
Embryo rescue
Abnormal embryo rescue seedlings
Normal grape seedlings
Acclimation and transplantation
