橡胶树不同发育时期花药愈伤生长发育对低温的响应  

Response of Growth and Development of Anther Callus in Different Developmental Stages to Low Temperature in Hevea brasiliensis

作　　者：王梦澳 李娜 吴日智 周权男[2] 彭素娜[2] 杨先锋[2] 黄天带[2] 曾长英 WANG Mengao;LI Na;WU Rizhi;ZHOU Quannan;PENG Suna;YANG Xianfeng;HUANG Tiandai;ZENG Changying(School of Tropical Agriculture and Forestry,Hainan University,Haikou,Hainan 570228,China;Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences/Haikou Key Laboratory of Tropical Plant Seedling Innovation/Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture and Rural Affairs/Hainan Key Laboratory for Cultivation&Physiology of Tropical Crops/State Key Laboratory Incubation Base for Cultivation and Physiology of Tropical Crops,Haikou,Hainan 571101,China;College of Plant Science and Technology,Huazhong Agricultural University,Wuhan,Hubei 430070,China)

机构地区：[1]海南大学热带农林学院,海南海口570228 [2]中国热带农业科学院橡胶研究所/海口市热带植物种苗创新重点实验室/农业农村部橡胶树生物学与遗传资源利用重点实验室/海南省热带作物栽培生理学重点实验室/省部共建国家重点实验室培育基地,海南海口571101 [3]华中农业大学植物科学技术学院,湖北武汉430070

出　　处：《热带作物学报》2024年第9期1877-1885,共9页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.32271915);国家天然橡胶产业技术体系项目(No.CARS33);中央级公益性科研院所基本科研业务费专项(No.1630022022001)。

摘　　要：橡胶树花药再生体系是我国最早建立的橡胶树体胚植株再生技术体系,其产生的再生植株为自根幼态无性系,兼具实生苗生长迅速、抗逆性强和芽接苗遗传同质的优点,同时因发育时期从老态恢复至幼态,产量提升20%以上。然而每年花蕾采集期约为60d,无法全年供应由花药壁细胞脱分化形成的愈伤组织,造成依托于该材料的研究无法持续进行。本研究以橡胶树品种热研73397雄蕊为材料,研究不同发育时期的花药愈伤(0、7、14、21、28、35、42 d)在15、20℃两个低温下保存15 d其愈伤诱导和体细胞胚发生发育的情况。结果表明:15℃和20℃两个温度对低温保存后愈伤诱导、体细胞胚发生各阶段除胚性愈伤数显著低于对照组外,其余无显著影响;发育时期对低温保存后愈伤诱导、体细胞胚发生各阶段均有显著影响(P<0.05),对子叶胚形成阶段影响最大,其中21d和35d花药愈伤组织经低温保存后其子叶胚数量分别显著高于和低于对照组子叶胚(P<0.05);15℃和20℃低温保存15 d后不同发育时期花药愈伤组织与对照组相比,可延长花药初代愈伤供应时间15~60 d,初代子叶胚供应时间27~95 d。最优方案为发育时期21 d的花药愈伤在15℃低温下保存15 d,可延长花药愈伤和初代子叶胚供应60 d和60~95 d,该处理虽降低了46.61%愈伤数,但促进了子叶胚形成,子叶胚数提高了27.18%。本研究实现了橡胶树花药愈伤的短期保存,有效解决了因花期短导致遗传转化受体来源不足的困难,全年3个花季可开展实验时间由60 d延长至240 d,有助于加快橡胶树花药愈伤遗传转化体系优化研究。The anther regeneration system is the earliest in vitro tissue culture regeneration plant system of Hevea brasiliensis established in China,and the regenerated plants were named self-rooted juvenile clone,which have the advantages of rapid growth,high resistance as seedlings and genetic homogeneity as grafted seedlings,and yield increase by 1878热带作物学报第45卷more than 20%due to rejuvenation.However,the buds collection period is only about 60 d per year,which does not allow for a year-round supply of callus formed by dedifferentiation anther wall cells,resulting in unsustainable research relying on this material.In this study,Reyan73397 stamens were utilized to study the callus induction and somatic embryogenesis of anther callus at different developmental periods(0 d,7 d,14 d,21 d,28 d,35 d and 42 d),which were stored at two low temperatures of 15 and 20 for 15 d.The results sh℃℃owed that temperature 15℃and 20℃had no significant effect on the stages of callus induction and somatic embryogenesis after preservation,except that the number of embryogenic callus was significantly lower than that of the control the developmental period had significant effect on callus induction and somatic embryogenesis after preservation,and showed the greatest effect on the stage of cotyledonary embryo formation,of which the anther callus at 21 d and 35 d responded strongly,the number of cotyledonary embryos of 21 d anther callus was significantly higher than that of the control,and the number of cotyledonary embryos of 35 d anther callus was significantly lower than that of the control after preservation;the preservation of anther callus with different developmental periods for 15 d at 15℃and 20℃could extend the supply time of anther callus for 15~60 d,and primary cotyledonary embryos for 27-95 d compared to the control;the optimal solution was to keep the anther callus with developmental periods at 21 d at 15℃and 20℃for 15days,which could prolong the supply of anther callus and primary cotyledonary embry

关 键 词：橡胶树 遗传转化 花药愈伤 低温保存 体细胞胚发生 

分 类 号：S794.1[农业科学—林木遗传育种]