作 者：
李昂;戴冬;赵立;韦壮敏;丁德宽;袁晓慧;叶俊丽;谢宗周;邓秀新;柴利军
单 位：
华中农业大学园艺林学学院·园艺植物生物学教育部重点实验室;城固县果业技术指导站;龙陵恒冠泰达农业发展有限公司
摘 要：
【目的】新资源获得是开展育种的前提和基础,通过两种砧木杂交,创造出聚合两种砧木优良性状的新型砧木资源材料。【方法】以枳雀为母本、枳为父本进行杂交实验,结合胚抢救技术获得子一代植株,利用形态学观察及SSR分子标记技术对子一代进行鉴定。【结果】三年胚抢救实验共接种3 146粒种子,获得胚抢救幼苗1250株,成苗率为39.73%。其中通过形态学鉴定出41株杂种苗,共筛选出8对SSR引物来对父本母本进行区分,其中的CS014RCS014F、CS065R-CS065F引物组合可以准确地鉴定出杂种后代,通过SSR分子标记鉴定出20株有性杂种植株。所获得的61株杂种资源为将来深入开展柑橘砧木育种研究工作奠定了材料基础。【结论】以多胚性柑橘为母本的杂交组合获得的成熟种子,难以通过直接播种的方式获得杂种苗。将胚拯救与分子鉴定相结合,通过SSR分析,找到可靠的SSR引物对鉴定杂交后代,为柑橘砧木育种的进一步研究奠定了物质基础。
译 名：
Creation and identification hybrid population from Zhique × Trifoliate orange based on embryo rescue technology
作 者：
LI Ang;DAI Dong;ZHAO Li;WEI Zhuangmin;DING Dekuan;YUAN Xiaohui;YE Junli;XIE Zongzhou;DENG Xiuxin;CHAI Lijun;Key Laboratory of Horticultural Plant Biology, Ministry of Education/Key Laboratory of Horticultural Crop Biology and Genetic Improvement (Central Region), MOA, Huazhong Agricultural University;Chenggu Fruit Industry Technical Guidance Station;Longling Hengguan Taida Agricultural Development Co. LTD;
关键词：
Citrus;;Zhique;;Trifoliate orange;;Embryo rescue;;Hybrid population
摘 要：
【Objective】The research and utilization of resistant rootstock resources of Citrus in China started relatively late, and the breeding rootstock is still relatively delayed in comparison with that in the developed countries. In order to obtain hybrid offspring population of Zhique( Citrus wilsonii Tanaka) Raf.) and Trifoliate orange[Poncirus trifoliata(L.) Raf], we conducted this study. Trifoliate orange is used as a kind of citrus rootstock widely in China. It has good affinity and general resistance. But the major defect is the insufficiency of iron absorption. Zhique is a kind of local citrus rootstock resource reported in Chenggu county, Shanxi province recently. It has good ability in using iron. In order to fully combine the beneficial properties, we carried out hybridization combined with embryo rescue technique to obtain hybrid offspring.【Methods】The species Zhique was planted in Chenggu county, Shanxi province. In this experiment, the Zhique's plants, leaves, flowers, fruits and other materials used for morphological description were all collected from a tree which was estimated more than 40 years old. Zhique was used as female parent and Trifoliate orange was used as male parent. The pollen of Trifoliate orange was collected in HZAU Germplasm Resources Nursery during morning. After the collection, the pollen was dried in dryer with no light at 27-29 ℃. We stored the pollen in 1.5 mL centrifuge tubes at4 ℃ when they were fully dried. We used acetic acid magenta to test the activity before the pollination and bagged the flowers after pollination. The hybrid fruits were picked 80-85 days after the pollination.The fruits were stored in 4 ℃ refrigerator before the embryo rescue was done. The embryos were inoculated on the shooting medium(MT+KT 0.5 mg · L~(-1)+BA 0.5 mg · L~(-1)+NAA 0.5 mg · L~(-1)) under illumination culture until the bud grew up. The shoots were moved on rooting medium(l/2 MT+IBA 0.1 mg · L~(-1)+NAA 0.5 mg · L~(-1)+Activated Carbon 0.5 mg · L~(-1)) until the root grew up. After subculture the plants were transfered into water for 1 month adaptation. Finally we put the well adapted seedlings into small pots with nutritive soil, and moved the pots in greenhouse. There were 1 133 progeny seedlings survival in the end, some of them were identified with morphology of the male parent, but the rest had to be identified by molecular markers. 8 of the primer pairs of the 10 possible SSR primer pairs were able to be used for the identification of the true hybrid seedlings.【Results】61 hybrid offspring were obtained.Among them 41 of the hybrid plants were identified with the trifoliate morphologic characteristics. Another 20 hybrid plants were identified by SSR molecular marker primer pair CS014 R-CS014 F and CS065 R-CS065 F.he hybrid rate was 4.1%.【Conclusion】Hybrid offsprings derived from the cross between Zhique and Trifoliate orange were successfully identified by means of morphology and SSR marker selection from the seedlings obtained with embryo rescue which created the materials for the further study of citrus rootstock breeding.
