Character Identification and Genetic Analysis of Progeny from Distant Hybrid Between Gossypium hirsutum and G. thurberi

doi:10.13560/j.cnki.biotech.bull.1985.2023-0539

Abstract

Abstract:

Distant hybridization and polyploidization are fundamental processes for the development of new species and germplasm innovation. In this study, we obtained fertile offsprings by interspecific distant hybridization and chromosome doubling between Gossypium hirsutum and Gossypium thurberi. The objective is to penetrate beneficial agronomic traits of G. thurberi into G. hirsutum and expand the genetic diversity of G. hirsutum. In this study, we obtained hybrid F₁ by synthesizing the female parent G. hirsutum with the male parent G. thurberi, and had allohexaploid cotton by chromosome doubling. And then we examined the morphology, cytology, physiology, biochemistry and SSR molecular markers of the F₁ and allohexaploid. The results of morphological identification showed that the allohexaploid plant was similar to the heterotriploid, with mid-parents'characters. The allohexaploid had more stamens than the parent and F₁had, with reddish spots, which were different from the parents and F₁. Cytological identification showed that stomatal density was in a decreasing trend with the increase of ploidy; while stomatal length, chloroplast number and pollen grain diameter all showed an increasing trend. The meiotic behavior of heterotriploids and allohexaploids revealed that both normal and abnormal tendencies, with more polyploids in triploids and significantly more normal tetrads in hexaploids. In triploids, the percentage of normal tetrads was 24.33%, while the percentage of normal tetrads was 82.17% in the polyploid hexaploids, indicating the recovery of hexaploid fertility. Physiological and biochemical results showed that the activities of enzymes increased with the increase of ploidy in different generations. The SSR molecular marker identification results revealed that distant hybridization led to chromosomal recombination in triploid hybrids and heterohexaploids, resulting in the amplification of both parental bands and new specific bands. The comprehensive results show that hybrids are successfully synthesized by distant hybridization and fertile heterohexaploids are successfully obtained by chromosome doubling, which provides valuable materials for cotton germplasm innovation and further genetic research.

Key words: G. thurberi, distant hybridization, allohexaploid, character identification, physiological and biochemical index, cytology identification, SSR

HOU Lin-hui, ZHENG Yun, RONG Er-hua, WU Yu-xiang. Character Identification and Genetic Analysis of Progeny from Distant Hybrid Between Gossypium hirsutum and G. thurberi[J]. Biotechnology Bulletin, 2023, 39(12): 128-135.

Figures/Tables 13

References 25

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类名 Classific name	染色体组 Genome	染色体数Number of chromosomes	来源 Source
陆地棉 G. hirsutum	AD₁	2n=4X=52	中国农科院棉花研究所 Cotton Research Institute, CAAS
瑟伯氏棉 G. thurberi	D₁	2n=2X=26	国家种质三亚野生棉圃 Sanya National Germplasm Wild Cotton Garden

类名 Classific name	染色体组 Genome	染色体数Number of chromosomes	来源 Source
陆地棉 G. hirsutum	AD₁	2n=4X=52	中国农科院棉花研究所 Cotton Research Institute, CAAS
瑟伯氏棉 G. thurberi	D₁	2n=2X=26	国家种质三亚野生棉圃 Sanya National Germplasm Wild Cotton Garden

正向引物Forward primer	引物序列Primer sequence（5'-3'）	反向引物Reverse primer	引物序列Primer sequence（5'-3'）
BNL4108-F	TCCACCATTCCCGTAAATGT	BNL4108-R	TGGCCAAGTCATTAGGCTTT
BNL4053-F	TGAAGGCTTTGAAGCAAACA	BNL4053-R	AAGCAAGCACCAAGTTAGCC
NAU2026-F	GAATCTCGAAAACCCCATCT	NAU2026-R	ATTTGGAAGCGAAGTACCAG
NAU1355-F	ATCTGTTTACGCCACTCTCC	NAU1355-R	CCAGCCTTTGACATTTTTCT
NAU1169-F	GGGTAGTAGCTTTTATGATAGGG	NAU1169-R	CCATTCCTTCCCCTAATTCT
NAU1157-F	GAGTTTGGTTCTGGGTTGAG	NAU1157-R	GATCCTTTTCATCTCCTCCA
NAU1052-F	CGCAGATAAAGGATGGATTT	NAU1052-R	AGAGCTGGAGGACATAACAAA
NAU1042-F	CATGCAAATCCATGCTAGAG	NAU1042-R	GGTTTCTTTGGTGGTGAAAC
NAU1164-F	CCAACGCTAATTCTACCTCCT	NAU1164-R	GCGGGTAATTGTAGTACATGC
NBRI_GI1015-F	GAGTTTGGCTCCGAGTTGAG	NBRI_GI1015-R	GCGAGTGAACTCACCGATAA
NBRI_GM7-F	GTGATACCTGGAAAGATGTGA	NBRI_GM7-R	TCCTGATGCTGTTTTAGTCTT
NBRI_Gpd_21-F	GCAATTGTGGAAGAGAAGTAA	NBRI_Gpd_21-R	GGACCTCTGAGTCCATTTATT
NBRI_Gpd_85-F	CTCAAAAACCCCTTTAGTCTC	NBRI_Gpd_85-R	GAGTGAAAACCCGCAAAG

正向引物Forward primer	引物序列Primer sequence（5'-3'）	反向引物Reverse primer	引物序列Primer sequence（5'-3'）
BNL4108-F	TCCACCATTCCCGTAAATGT	BNL4108-R	TGGCCAAGTCATTAGGCTTT
BNL4053-F	TGAAGGCTTTGAAGCAAACA	BNL4053-R	AAGCAAGCACCAAGTTAGCC
NAU2026-F	GAATCTCGAAAACCCCATCT	NAU2026-R	ATTTGGAAGCGAAGTACCAG
NAU1355-F	ATCTGTTTACGCCACTCTCC	NAU1355-R	CCAGCCTTTGACATTTTTCT
NAU1169-F	GGGTAGTAGCTTTTATGATAGGG	NAU1169-R	CCATTCCTTCCCCTAATTCT
NAU1157-F	GAGTTTGGTTCTGGGTTGAG	NAU1157-R	GATCCTTTTCATCTCCTCCA
NAU1052-F	CGCAGATAAAGGATGGATTT	NAU1052-R	AGAGCTGGAGGACATAACAAA
NAU1042-F	CATGCAAATCCATGCTAGAG	NAU1042-R	GGTTTCTTTGGTGGTGAAAC
NAU1164-F	CCAACGCTAATTCTACCTCCT	NAU1164-R	GCGGGTAATTGTAGTACATGC
NBRI_GI1015-F	GAGTTTGGCTCCGAGTTGAG	NBRI_GI1015-R	GCGAGTGAACTCACCGATAA
NBRI_GM7-F	GTGATACCTGGAAAGATGTGA	NBRI_GM7-R	TCCTGATGCTGTTTTAGTCTT
NBRI_Gpd_21-F	GCAATTGTGGAAGAGAAGTAA	NBRI_Gpd_21-R	GGACCTCTGAGTCCATTTATT
NBRI_Gpd_85-F	CTCAAAAACCCCTTTAGTCTC	NBRI_Gpd_85-R	GAGTGAAAACCCGCAAAG

倍性 Ploidy	单分体 Monad	二分体 Dyad	三分体 Triad	四分体 Tetrad	多分体 Polyad	总数 Total
F₁(3X)	73(12.17)	25(4.17)	38(6.33)	146(24.33)	318(53.00)	600
M₁(6X)	63(10.50)	15(2.50)	17(2.83)	493(82.17)	12(2.00)	600