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Production and characterization of asymmetric hybrids between upland cotton Coker 201 (Gossypium hirsutum) and wild cotton (G. klozschianum Anderss)

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Abstract

Asymmetric somatic hybrids were obtained between Gossypium hirsutum Coker 201 and wild cotton G. klozschianum Anderss. An investigation on the effect of ultraviolet (UV) irradiation on donor protoplasts was carried out, and the lethal dose was determined to be 38.7 J cm−2. We firstly screened the putative hybrids by the color of the calli produced, followed by morphological, cytological, and molecular analysis of putative hybrid plants. Most regenerated plants derived from fused protoplasts displayed a recipient-like morphology, while some showed an intermediate phenotype between Coker 201 and G. klozschianum. Chromosome numbers in these somatic hybrids ranged from 54 to 74. The hybrids were verified by random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR). Absence or co-existence of parents’ genome DNA fragments was identified through molecular analysis. The heredity of cytoplasm was investigated by cleaved amplified polymorphic sequence (CAPS) analysis using mitochondrial and chloroplast universal primer pairs. The results indicated that recombination and rearrangements might have occurred in some regions of mitochondria (mt) and chloroplast (cp) DNA. To our knowledge, this is the first report about asymmetric protoplast fusion in cotton, and the hybrids obtained would be useful for breeding programs.

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Abbreviations

CAPS:

Cleaved amplified polymorphic sequence

cpDNA:

Chloroplast DNA

CPW:

Cell and protoplast washing solution (Frearson et al. 1973)

CTAB:

Cetyltrimethyl ammoniumbromide

FDA:

Fluorescein diacetate

GFP:

Green fluorescent protein

IBA:

Indole-3-butyric acid

IOA:

Iodoacetamide

MES:

2, (N-morpholino) ethane sulfonic acid

MSB:

MS medium (Murashige and Skoog 1962) and B5 (Gamborg et al. 1968) vitamins

mtDNA:

Mitochondrial DNA

PAGE:

Polyacrylamide gel electrophoresis

RAPD:

Random amplified polymorphic DNA

SSR:

Simple sequence repeat

UV:

Ultraviolet

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Acknowledgments

The author X. Y. Yang thanks Y. X. Zhang and L. L. Tu for technical assistance with the molecular analysis. This research was supported by program for New Century Excellent Talents in University (NCET-04-0739) and National Basic Research program of China (2004CB117301).

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  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei, 430070, P.R. China

    Xi-yan Yang, Xian-long Zhang, Shuang-xia Jin, Li-li Fu & Ling-gang Wang

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  1. Xi-yan Yang
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  2. Xian-long Zhang
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Correspondence to Xian-long Zhang.

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Yang, Xy., Zhang, Xl., Jin, Sx. et al. Production and characterization of asymmetric hybrids between upland cotton Coker 201 (Gossypium hirsutum) and wild cotton (G. klozschianum Anderss). Plant Cell Tiss Organ Cult 89, 225–235 (2007). https://doi.org/10.1007/s11240-007-9245-0

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