Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (6): 968-980.doi: 10.3864/j.issn.0578-1752.2019.06.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Effects of Different Red and Blue Ratios on the Somatic Embryogenesis and Plant Regeneration of Cotton

WEI Xi1,2,WANG QianHua2,GE XiaoYang2,CHEN YanLi2,DING YanPeng2,ZHAO MingZhe1(),LI FuGuang2()   

  1. 1 College of Agricultural, Shenyang Agricultural University, Shenyang 110866
    2 Institute of Cotton Research, Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology, Anyang 45000, Henan
  • Received:2018-11-29 Accepted:2019-01-27 Online:2019-03-16 Published:2019-03-22
  • Contact: MingZhe ZHAO,FuGuang LI E-mail:soyshen@126.com;aylifug@163.com

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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

Table 1

Compositions of culture medium at somatic embryogenesis stage of cotton"

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

Fig. 1

Cotton somatic embryogenesis A: Flow chart of cotton somatic embryogenesis; B: 5-7 mm cotton hypocotyl segments; C: Early callus; D: Late callus; E: Embryogenic callus; F: Globular embryo; G: Heart-shaped embryo; H: Torpedo embryo; I: Cotyledon embryo; J: Regenerated cotton plant"

Fig. 2

Effect of different light quality treatments on CPR A: Callus phenotype under four different red and blue ratios; B: Callus proliferation rate under four different light quality treatments was carried out by the least significant difference (LSD) method (P < 0.05). Different lower-case letters in B denote statistical differences of pairwise comparisons among each group of bars according to LSD test (P<0.05). The same as below"

Fig. 3

Effects of different light quality treatments on undifferentiated callus A: Phenotype of undifferentiated callus under four different red-blue ratios; B: Statistical analysis of the proliferation of undifferentiated callus under four different light quality treatments "

Fig. 4

Effects of different light quality treatments on embryogenic callus induction in cotton A: Phenotypes of hypocotylic callus with four different red and blue ratios; B: Statistical analysis of differentiation rate of embryonic callus with four different light quality treatments. ECs: Embryogenic callus"

Fig. 5

Effects of different light quality treatments on the induction stage of cotton embryoids A: The phenotype of somatic embryos under four different red and blue ratios; B: The statistics of somatic embryos under four different light quality treatments"

Fig. 6

Effects of different light quality treatments on the number of hypocotyls from SEs"

Fig. 7

Effects of different light quality treatments on rooting of cotton hypocotyls A: Phenotypes of rooting under four different ratios of red and blue; B: Statistics of rooting rates under four different light quality treatments"

Fig. 8

Effects of different light qualities on cotton seedling regeneration and plant height A: The phenotype of regenerated cotton plants under four different red-blue ratios; B: the statistics of seedling number of hypocotyl under four different light quality treatments; C: the statistics of seedling number of hypocotyl under four different light quality treatments"

Fig. 9

Effects of different light qualities on chlorophyll concentration of cotton regenerated seedlings A: Leaf phenotype of regenerated cotton under four different red-blue ratios; B: Statistical analysis of chlorophyll concentration under four different light quality treatments "

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