高效草菇分子标记辅助杂交育种方法的建立与应用

Establishment and application of an efficient cross breeding method assisted by molecular markers of Volvariella volvacea

摘要:

【目的】建立一种高效草菇分子标记辅助杂交育种方法，并将其应用于选育低温高产草菇的单孢杂交中。【方法】通过出菇试验筛选获得高产草菇菌株，与低温出菇菌株VH3一同作为杂交亲本；采用交配型基因分子标记区分草菇单孢子的交配型，并完成杂交配对工作；最后结合快速筛选耐低温草菇菌种技术与杂交子真实性的鉴定方法，在栽培出菇试验前剔除部分不耐低温的草菇杂交子。【结果】出菇试验表明，屏优1号的生物转化率最高，用作杂交育种的高产亲本菌株。单孢杂交配对后，最初的496株草菇可能杂交菌株经初筛后，只剩余172株较耐低温的杂交菌株，使后期出菇试验的工作量减少了65%；杂交子出菇试验表明，在28 °C出菇温度下，VV093杂交菌株的生物转化率显著高于两个亲本菌株，且农艺性状较好。【结论】建立了一种高效草菇分子标记辅助杂交育种的方法，包括亲本菌株的筛选、单孢交配型的区分、单孢杂交、杂交子的鉴定和筛选等，并在低温高产草菇单孢杂交育种中成功应用。

Abstract:

[Objective] In order to establish an efficient cross breeding method assisted by molecular markers of Volvariella volvacea and apply it to monospore cross breeding for the cold-tolerant and high-yield Volvariella volvacea strain. [Methods] In this study, the high-yield strain Pingyou No. 1 tested by cultivation experiment and the cold-tolerant strain VH3 were chosen as crossing parent strains. Then used the molecular marker of mating-type genes in Volvariella volvacea to verify mating-type of Volvariella volvacea monospore strain and completed the cross breeding. At last part of the cold-intolerant hybrids were eliminated before cultivation experiment using techniques of fast screening of the cold-tolerant Volvariella volvacea strain and identification of the hybrid strains. [Results] The result of cultivation experiment showed that the strain Pingyou No. 1 got the highest biological efficiency, which would be chosen as the high-yield parent strain. The 172 cold-tolerant hybrid strains were screened out from the original 496 possible hybrid strains so that the workload of cultivation experiment in the late would get a 65% reduction. The result of hybrid strains cultivation experiment showed that VV093 got higher biological efficiency than Pingyou No. 1 or VH3 at 28 °C. [Conclusion] Establishment of an efficient cross breeding method assisted by molecular markers of Volvariella volvacea included selection of parental strains, identification of monospore strains’ mating-type genes, monospore cross breeding, screening and identification of possible hybrid strains, and it was applied to monospore cross breeding for the cold-tolerant and high-yield Volvariella volvacea strain successfully.