不同红蓝配比的光质对棉花体细胞胚胎发生 和植株再生的影响

doi:10.3864/j.issn.0578-1752.2019.06.002

摘要/Abstract

摘要：

【目的】设置不同的光质组合,明确不同光质对棉花体细胞胚胎发生的影响,为提高棉花体细胞胚胎发生能力和加快棉花转基因过程提供依据。【方法】分别设置不同红蓝配比的光质组合B﹕R（蓝﹕红）=1﹕1、B﹕R（蓝﹕红）=3﹕1、B﹕R（蓝﹕红）=1﹕3和DL（日光灯）,将CCRI12（中棉所12）棉花下胚轴切段依次在愈伤诱导培养基上（callus induction medium,CIM）、胚性愈伤组织诱导培养基上（embryogenic callus differentiation medium,ECDM）、体细胞胚诱导培养基上（embryoid induction medium,EIM）和再生植株诱导培养基（regenerated-plantlet induced medium,RIM）上培养。统计在4种不同光质配比下,体细胞胚胎发生各个阶段对应的指标,包括愈伤增殖率（callus proliferation rate,CPR）、愈伤增殖量、胚性愈伤分化率、体细胞胚数量、生根率、下胚轴数量、再生棉花植株数量、叶绿素浓度等。【结果】与B﹕R=1﹕3和DL光质组合相比,B﹕R=1﹕1和B﹕R=3﹕1的光质组合均显著促进了愈伤的增殖率,但抑制体细胞胚长出下胚轴;与B﹕R=1﹕1和B﹕R=3﹕1相比,B﹕R=1﹕3和DL光质组合均促进胚性愈伤分化和诱导体细胞胚的产生,在诱导下胚轴生根方面,B﹕R=1﹕1组合生根率明显高于其他3种处理;B﹕R=1﹕3和B﹕R=1﹕1处理后下胚轴成苗数、植株高度和叶绿素浓度明显高于B﹕R=3﹕1和DL光质组合,B﹕R=1﹕3的促进效果最明显。【结论】明确了不同光质组合在体细胞胚胎发生各个阶段的调控作用,在体细胞胚胎发生各个阶段可以选择性地应用最优的光质组合。其中红蓝配比为B﹕R=1﹕3的光质组合在整体上能促进棉花体细胞胚胎的发生。

关键词: 陆地棉, 红蓝光配比, 体细胞胚胎发生, 愈伤, 胚性愈伤, 体细胞胚, 再生苗

Abstract:

【Objective】Different light quality combinations were set up to clarify the effects of different light quality on cotton somatic embryogenesis, and to provide basis for improving cotton somatic embryogenesis ability and accelerating cotton genetically modified process. 【Method】The light quality combinations B﹕R (blue﹕red) = 1﹕1, B﹕R (blue﹕red) = 3﹕1, B﹕R (blue﹕red) = 1﹕3 and DL (fluorescent lamp) with different red and blue ratios were set respectively. The hypocotyl segments of CCRI12 cotton were cultured on the callus induction medium (CIM), embryogenic callus induction medium (ECDM), somatic embryo induction medium (EIM) and root induction medium (RIM) in turn. Under four different light quality ratios, the corresponding indicators of somatic embryogenesis at different stages were counted, including callus proliferation rate (CPR), callus proliferation, embryogenic callus differentiation rate, somatic embryo number, rooting rate, hypocotyl number, regenerated cotton plant number, chlorophyll concentration, etc.【Result】Compared with B﹕R=1﹕3 and DL light quality combinations, B﹕R=1﹕1 and B:R=3﹕1 light quality combinations significantly promoted callus proliferation. Compared with B﹕R=1﹕1 and B﹕R=3﹕1 light quality combinations, B﹕R=1﹕3 light quality combination significantly increased weight proliferation of callus without EC differentiation ability. B﹕R=1﹕3 and DL light quality combinations had significantly higher embryonic callus differentiation rate than B﹕R=1﹕1 and B﹕R=3﹕1 light quality combinations. The number of somatic embryos induced by B﹕R=1﹕3 treatment was significantly higher than that of B﹕R=1﹕1 and B:R=3﹕1 light quality combination, but there was no significant difference compared with DL; the number of hypocotyls induced by B﹕R=1﹕1 and B﹕R=3﹕1 light quality combination was significantly lower than that induced by B﹕R=1﹕3 and DL light quality combination, and the number of somatic embryos induced by B﹕R=1﹕1 light quality combination and B﹕R=3﹕1 light quality combination was significantly lower than that induced by B﹕R=1 light quality combination and B﹕R=3﹕1 light. The plant height of B﹕R=1﹕1 and B﹕R=1﹕3 combinations was significantly higher than that of B﹕R=1﹕3 and DL combinations; the chlorophyll content of B﹕R=1﹕3 treatment was higher than that of B﹕R=1﹕1 treatment, but the chlorophyll content of B﹕R=1﹕3 treatment was significantly higher than that of B﹕R=3﹕1 and DL light quality combination.【Conclusion】The regulation of different light quality combinations in different stages of somatic embryogenesis was clarified. Optimal light quality combinations could be selectively applied in different stages of somatic embryogenesis. The combination of red and blue B﹕R=1﹕3 can promote the somatic embryogenesis of cotton.

Key words: Gossypium hirsutum, red and blue ratio, somatic embryogenesis, callus, embryogenic callus, somatic embryo, regenerated seedlings

魏喜,王倩华,葛晓阳,陈艳丽,丁颜朋,赵明哲,李付广. 不同红蓝配比的光质对棉花体细胞胚胎发生和植株再生的影响[J]. 中国农业科学, 2019, 52(6): 968-980.

WEI Xi,WANG QianHua,GE XiaoYang,CHEN YanLi,DING YanPeng,ZHAO MingZhe,LI FuGuang. Effects of Different Red and Blue Ratios on the Somatic Embryogenesis and Plant Regeneration of Cotton[J]. Scientia Agricultura Sinica, 2019, 52(6): 968-980.

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名称 Name	基础培养基 MSB (mL·L^-1)	蔗糖 Sucrose (g·L^-1)	葡萄糖 Glucose (g·L^-1)	6-糠氨基嘌呤 Kinetin (KT) (mg·L^-1)	2,4-二氯苯氧乙酸 2,4-D (mg·L^-1)	吲哚乙酸 IAA (mg·L^-1)	6-苄氨基腺嘌呤 6-BA (mg·L^-1)	琼脂 Gelose (g·L^-1)
愈伤诱导培养基CIM	65		28	0.1	0.1			2.2
胚性愈伤诱导培养基ECDM	65		28	0.15		0.05		2.2
体细胞胚诱导培养基EIM	65	28		0.1			0.2	2.2
再生苗诱导培养基RIM	65	25				0.05	0.05