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作 者:李爽 金峰 向旭[1] LI Shuang;JIN Feng;XIANG Xu(Institute of Fruit Tree Research,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China)
机构地区:[1]广东省农业科学院果树研究所,广东广州510640
出 处:《广东农业科学》2021年第2期41-49,共9页Guangdong Agricultural Sciences
基 金:广东省农业科学院院长基金(201816);广东省农业科学院青年导师制项目(R2018QD-025);广东省重点领域研发计划项目(2018B020202011);广东省农业科学院学科团队项目(201626TD)。
摘 要:【目的】开展荔枝(Litchi chinensis)叶片胚性愈伤组织诱导和增殖研究,为荔枝再生体系技术的建立及应用打下基础。【方法】以荔枝叶片为试验材料对胚性愈伤组织诱导的影响要素(基因型、材料来源、培养基成分等)进行研究,并开展胚性愈伤组织继代增值试验。【结果】将萌发后2~4周的无菌叶片切成1 cm^(2)大小并沿垂直于主叶脉方向切开2道伤口,正面朝下接种于培养基MS+2,4-D 1.5 mg/L+KT 2.0 mg/L+蔗糖30 g/L+琼脂7 g/L+PVP 500 mg/L+肌醇100 mg/L上,经2~3次继代后,三月红和御金球品种能诱导出状态较好、可用于后续试验的胚性愈伤组织,诱导率分别为21.67%和42.50%。在添加2,4-D 1.5 mg/L+NAA 2.0 mg/L的培养基上继代增殖率最高达364%,且在植物生长调节剂减半的继代培养基上交替培养生长效果最好。【结论】三月红和御金球品种能诱导出胚性愈伤组织,添加肌醇100 mg/L+2,4-D 1.5 mg/L+KT 2.0 mg/L的组合有利于胚性愈伤组织发生,2,4-D 1.5 mg/L+NAA 2.0 mg/L组合有利于胚性愈伤组织的增殖培养,且愈伤组织在该组合减半并添加2.0 mg/L AgNO3的培养基上交替培养效果最好。【Objective】The induction of embryogenetic callus and proliferation of litchi leaves were studied in order to lay a good foundation for the establishment and application of litchi regeneration system technology.【Method】With the litchi leaves as materials,the influencing effects(genotype,source of materials,components of culture medium,etc.)of litchi leaves on embryogenic callus induction were studied,and the proliferation experiment on subculture proliferation of embryogenic callus was carried out.【Result】The sterile leaves germinating for about 2-4 weeks were cut into a size of 1 cm^(2) with two cuts perpendicular to the main vein,and were inoculated facing down to the medium of MS+2,4-D 1.5 mg/L+KT 2.0 mg/L+sucrose 30 g/L+agar 7 g/L+PVP 500 mg/L+inositol 100 mg/L.After 2-3 subcultures,the varieties of‘Sanyuehong’and‘Yujinqiu’could induce embryogenic callus with better state and these callus could be used in subsequent experiments,with the induction rates of 21.67%and 42.50%,respectively.The subculture proliferation rate on the 2,4-D 1.5 mg/L+NAA 2.0 mg/L medium was up to 364%,and the growth effect of embryogenic callus was the best when it was alternately cultured on the subculture medium with half the plant growth regulator.【Conclusion】Embryogenic callus could be induced from‘Sanyuehong’and‘Yujinqiu.’The combination of inositol 100 mg/L+2,4-D 1.5 mg/L+KT 2.0 mg/L was beneficial to embryogenic callus formation.The plant growth regulator combination of 2,4-D 1.5 mg/L+NAA 2.0 mg/L was beneficial to the proliferation culture of embryogenic callus,and the growth effect of embryogenic callus was the best when it was alternately cultured on the medium with half of the plant growth regulator and 2.0 mg/L AgNO3.
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