分子标记辅助甜瓜抗蔓枯病基因的聚合及品种改良

doi:10.3864/j.issn.0578-1752.2015.03.12

Abstract

Abstract: 【Objective】Gummy stem blight (GSB) is caused by Didymella bryoniae and is a serious fungal disease of melon (Cucumis melon L.). The resistance of melon varieties carrying a single resistance gene is not enough to the disease because of the isolates variation of Didymella bryoniae. Therefore, this study aims to develop a molecular marker-assisted selection system and provide an important intermediate materials for melon disease-resistance breeding. 【Method】Firstly, the single resistant sources PI140471, PI157082, PI511890, PI482398 and PI420145 were used to obtain polymerization resistant sources (082-471, 082-890, 082-398, 145-471, 145-082, 145-890, 145-398 and 890-398). Secondly, three different gradient spores vaccination identification (5×10⁵ spores/mL, 5×10⁷ spores/mL and 5×10⁹ spores/mL) was employed to distinguish the resistance of F₁ plant of eight different polymerization resistant sources. The gene combinations of 145-471 and 145-398 with their resistance significantly improved were screened by inoculation identification results of spring and autumn. Single resistant sources PI140471, PI482398 and PI420145 were used as the donor parents with the gummy stem blight resistance genes Gsb-1 Gsb-4 and Gsb-6, respectively. Commercial melon cultivar Baipicui was used as the receipt parent to cross and backcross with the donor parents. Marker assisted selection (MAS) and gradient spores vaccination identification (5×10⁵ spores/mL, 5×10⁷ spores/mL and 5×10⁹ spores/mL) were used in each backcross and self-cross progeny. 【Result】The single resistant sources showed lower resistance than polymerization individuals and showed selective resistance to the different gummy stem blight spore concentrations (5×10⁵ spores/mL, 5×10⁷ spores/mL and 5×10⁹ spores/mL). The detection results of molecular markers showed that all the individuals of 145-471 (or 145-398) F₇ and BC₅F₄ generation had two resistance genesof Gsb-1 and Gsb-6 (or Gsb-4 and Gsb-6). The polymorphism of molecular markers was different among the parents. The PCR amplification of SSR marker CMCT505 could augment one specific fragments of 189 bp in single resistant source PI140471. The PCR amplification of SSR marker CMTA170a could augment one specific fragments of 121 bp in single resistant source PI482398. A 1 800 bp specific fragment was obtained with SCAR marker SGSB₁₈₀₀ in resistant source PI420145. The polymerization individual 145-471 could be amplified two fragments of 189 bp and 1 800 bp by SSR marker CMCT505 and SCAR marker SGSB₁₈₀₀. The polymerization individual 145-398 could be amplified two fragments of 121 bp and 1 800 bp by SSR marker CMTA170a and SCAR marker SGSB₁₈₀₀. At F₇ generation, the pyramiding lines with homozygosity at both gene loci were obtained. The polymerization individuals showed higher resistance than single resistant sources PI140471, PI482398 and PI420145. The individuals were resistant to melon gummy stem blight, which was accordant to the expected result of molecular detection. Improved Baipicui BC₅F₄ generation showed high resistance to melon gummy stem blight and had no significant differences in fruit weight, fruit shape index, fruit crispness, flesh texture, flesh thickness and soluble solids content compared with the Baipicui. A molecular marker-assisted selection system was developed and thus providing important intermediate materials for melon disease-resistance breeding. 【Conclusion】The results of disease resistance evaluation indicated that the molecular markers developed in the study were efficient in selecting three resistance genes by MAS. The molecular marker-assisted selection system developed in this study was efficient for pyramiding multiple resistant genes. The resistance improved materials of Baipicui would be used in breeding and further resistance gene pyramiding.

Key words: melon (Cucumis melon L.), marker-assisted selection, disease resistance, variety improvement

BI Yan-fei, XU Bing-hua, QIAN Chun-tao, GUO Jing, ZHANG Yong-bing, YI Hong-ping, CHEN Jin-feng. Pyramiding Disease Resistance Genes and Variety Improvement by Molecular Marker-Assisted Selection in Melon (Cucumis melon L.)[J].Scientia Agricultura Sinica, 2015, 48(3): 523-533.

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