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Plant Cell, Tissue and Organ Culture

, Volume 89, Issue 2–3, pp 225–235 | Cite as

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

  • Xi-yan Yang
  • Xian-long ZhangEmail author
  • Shuang-xia Jin
  • Li-li Fu
  • Ling-gang Wang
Original Paper

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.

Keywords

Asymmetric somatic hybrids Cotton Protoplast fusion UV irradiation 

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

Notes

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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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Xi-yan Yang
    • 1
  • Xian-long Zhang
    • 1
    Email author
  • Shuang-xia Jin
    • 1
  • Li-li Fu
    • 1
  • Ling-gang Wang
    • 1
  1. 1.National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhanP.R. China

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