陆地棉与拟似棉异源六倍体的合成与性状鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2022-1230

摘要/Abstract

摘要：

远缘杂交及多倍化是棉花种质创新的重要途径，通过陆地棉与野生拟似棉远缘杂交并加倍获得可育的杂种后代，为利用拟似棉中的优良基因拓宽陆地棉的遗传多样性提供理论依据和新材料。本研究以陆地棉为母本、拟似棉为父本杂交合成F₁并采用秋水仙素进行染色体加倍，获得异源六倍体植株，对其进行形态学（叶片表型）、细胞学（气孔密度、气孔长度、保卫细胞对中叶绿体数、花粉粒直径）、生理生化指标（SOD、POD、CAT、可溶性蛋白及叶绿素荧光参数指标）鉴定。通过流式细胞鉴定筛选出了3株六倍体植株。六倍体与三倍体及亲本性状统计结果显示，六倍体叶形指数与陆地棉无显著差异，说明六倍体叶形整体偏向陆地棉，而三倍体远缘杂种更多偏向父本拟似棉，六倍体叶面积及气孔长度增大、保卫细胞中叶绿体数显著增多；六倍体花的表型性状介于双亲之间，花粉粒直径大于三倍体呈极显著差异，六倍体正常花粉粒比为73.25%，三倍体正常花粉粒比为51.13%，说明六倍体育性在恢复；六倍体叶片中SOD、POD、CAT及可溶性蛋白含量显著大于其他世代；六倍体较其他世代最大荧光产量Fm差异显著，初始荧光Fo、叶片可变荧光Fv、原初光能转化效率（Fv/Fm）、PS II潜在光化学效率（Fv/Fo）以及开放的PS II捕获的激发能的效率（Fm/Fo）均表现出极显著差异。综合结果证明通过杂交加倍成功合成了陆地棉与拟似棉异源六倍体新种质，同时发现六倍体在抗逆性及光合作用表现优于三倍体。

关键词: 陆地棉, 拟似棉, 远缘杂交, 异源六倍体, 杂种致死, 性状鉴定, 生理生化指标

Abstract:

Distant hybridization and polyploidy is an important way for cotton germplasm innovation, and it may provide theoretical evidence and new materials by distantly hybridizing G. gossypioides with G. hirsutum and then chromosome doubling to use the excellent genes of G. gossypioides and to broaden the genetic diversity of G. hirsutum. In this study, we obtained allohexaploid cotton by synthesizing the female parent G. hirsutum with the male parent G. gossypioides, following with chromosome doubling by colchicine. And then we examined the morphology（leaf phenotype）, cytology（stomatal density, stomatal length, chloroplasts number in guard cell pairs, and pollen grain diameter）, and physiological and biochemical indexes（SOD, POD, CAT, soluble protein and chlorophyll fluorescence parameters of the hexaploid）. Three hexaploid plants were selected by flow cytometric identification. The statistical results of hexaploid, triploid and parental traits showed that there was not significant difference for the leaf shape index between hexaploid and G. hirsutum, indicating that the leaf shape of hexaploid was overall preferring to G. hirsutum, while the triploid distant hybrid was more inclined to paternal cotton G. gossypioides. The leaf area and stomatal length of hexaploid increased, and the number of chloroplasts in the guard cells significantly increased. The flower phenotypic traits of hexaploid were intermediate between two parents, but the pollen grain diameter of hexaploid was greater than the triploid, with the normal pollen grain ratio of 73.25% for hexaploid and of 51.13% for triploid, indicating the fertility recovering of hexaploid. The content of SOD, POD, CAT and soluble protein in hexaploid leaves were significantly greater than the other generations. There were significant differences in the initial fluorescence Fo, leaf variable fluorescence Fv, primary light energy conversion efficiency（Fv/Fm）, PS II potential photochemical efficiency（Fv/Fo）and open PS II capture excitation energy efficiency（Fm/Fo）between hexaploid and other generations. The overall results proved that the new germplasm of hexaploid between G. hirsutum and G. gossypioides were successfully synthesized by hybridization and chromosome doubling in our study, and it was tested that hexaploid has excellent performances superior to triploid especially in stress resistance and photosynthesis.

Key words: Gossypium hirsutum, Gossypium gossypioides, distant hybridization, allohexaploid, hybrid lethality, character identification, physiological and biochemical index

王一帆, 候林慧, 常永春, 杨亚杰, 陈天, 赵祝跃, 荣二花, 吴玉香. 陆地棉与拟似棉异源六倍体的合成与性状鉴定[J]. 生物技术通报, 2023, 39(5): 168-176.

WANG Yi-fan, HOU Lin-hui, CHANG Yong-chun, YANG Ya-jie, CHEN Tian, ZHAO Zhu-yue, RONG Er-hua, WU Yu-xiang. Synthesis and Character Identification of Allohexaploid Between Gossypium hirsutum and G. gossypioides[J]. Biotechnology Bulletin, 2023, 39(5): 168-176.

图/表 11

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类名Species	倍性Ploidy	染色体组Genome	染色体数Chromosome number	地理分布Geographical distribution
陆地棉 G. hirsutum	4X	（AD）₁	2n=4X=52	世界广泛栽培 Worldwide cultivation
拟似棉 G. gossypioides	2X	D₆	2n=2X=26	墨西哥瓦哈卡洲 Oaxaca, Mexico

类名Species	倍性Ploidy	染色体组Genome	染色体数Chromosome number	地理分布Geographical distribution
陆地棉 G. hirsutum	4X	（AD）₁	2n=4X=52	世界广泛栽培 Worldwide cultivation
拟似棉 G. gossypioides	2X	D₆	2n=2X=26	墨西哥瓦哈卡洲 Oaxaca, Mexico

材料 Material	平均荧光强度 Mean fluorescence intensity	倍性 Ploidy
P₁	229.85	4X
F₁	136.44	3X
P₂	80.91	2X
M₀-1	258.47	6X
M₀-2	264.87	6X
M₀-3	176.52	3X-6X
M₀-4	284.34	6X

材料 Material	平均荧光强度 Mean fluorescence intensity	倍性 Ploidy
P₁	229.85	4X
F₁	136.44	3X
P₂	80.91	2X
M₀-1	258.47	6X
M₀-2	264.87	6X
M₀-3	176.52	3X-6X
M₀-4	284.34	6X

材料Material	叶面积Leaf area/mm²	叶形指数Leaf index	叶裂数leaf lobes	表面是否有茸毛Surface fuzz	相对叶绿素含量SPAD
P₁	3 959.15±261.99 aA	0.69±0.08 cB	4-6	有	34.25±0.91 bBC
P₂	3 819.18±270.21 aA	1.47±0.19 aA	3-5	无	30.91±1.89 cC
F₁	1 666.25±113.43 cB	0.93±0.11 bB	4-6	无	35.03±1.08 bB
M₀	2 124.53±23.94 bB	0.69±0.06 cB	3-6	有	40.97±1.00 aA