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杏鲍菇杂交亲本的筛选及F1代菌株杂种优势的预测
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摘要
杂交育种是通过单核菌丝的杂交配对达到基因重组目的的方法,从世界各先进国家育种工作现状来看,杂交育种仍是育种中应用最广、效果最显著的一种方法。杏鲍菇是珍稀、高档的食用菌之一,具有很好的开发前景。目前,已广泛开展杏鲍菇的栽培研究,但育种较少,所以通过杂交育种获得优质、高产、适应性强的杏鲍菇是当今研究的工作重点。
     本试验以14个杏鲍菇菌株为试验材料,通过选择不同优良性状的亲本,进行单孢分离获得单核菌丝,然后进行配对杂交;通过遗传物质的重组,综合两亲本的优良性状,克服亲本原有的不良性状,从而育成符合人们需要的具有杂种优势的新品种。
     通过对14个杏鲍菇菌株菌丝生长情况,子实体农艺性状,营养成分,产量以及早熟性状的比较结果发现,产量较高的菌株为PL14,PL17,PL15;经济性状好的菌株是PL15,PL16,PL17和PL3;在24℃恒温下PL2和PL12菌株在母种培养基上生长速度比较快;耐高温的菌株为PL7,PL11,PL14;PL14菌株为早熟的品种。
     应用垂直板聚丙烯酰胺凝胶不连续电泳方法,对14个来源不同杏鲍菇菌株的菌丝体进行了酯酶同工酶检测,分析了菌株间的亲缘关系,并用聚类的方法对菌株间的遗传差异进行了研究。结果表明,各菌株在酶谱上有丰富的表达,且在Rf值0.672处都出现一条稳定一致的基本酶带,但在酶带条数、Rf值及酶活性上菌株之间存在着差异,个别菌株有特征的酶带,14个菌株的相似系数在0.333~1.000之间,存在着一定的遗传差异,但从总体来看它们之间的遗传变异并不丰富,从而说明现有的杏鲍菇种质资源比较狭窄,同时,也在一定程度上表明菌株的地理分布与遗传差异间并不存在必然的联系。
     通过采用平均连锁聚类分析方法,对14个杏鲍菇的相似系数进行聚类,结果发现,大部分是类间组合杂种优势较强,类内组合杂种优势较弱,因此,杂交亲本应选择不同类的菌株。通过单孢分离、配对杂交获得95个新组合,同样采用垂直板聚丙烯酰胺凝胶不连续电泳方法,对杂交亲本与杂交新组合进行酯酶同工酶分析,在杂交子代中出现22个“杂种新酶带”和“互补型酶带”,杂种优势较强,这样的组合有望选出优良品种。
The crossbreeding was a method to attain gene recombination by the combinations pairing of monokaryotic mycelium. Seen from the status quo of breeding in some developed countries, crossbreeding was one of the best effect methods in breeding and has been using most widely. P. eryngii was a kind of rare and advanced fungi which will have a development planning. Study on the p. eryngii cultivation has been performed more wide than crossbreeding, it is important to attain p. eryngii strains with properties of good quality, high-yield and vigorous adaptability.
    In this experiment, 14 p.eryngii strains were used as testing material. Monokaryotic mycelium was attainted by sing-spore isolation of strains which had different desirable characteristics, then they were paired. New varieties which coincided with people's need were attained by recombination of genetic material and generalization of desirable characteristics.
    The mycelium growth state, the agronomic characteristics of fruitbodies, nutritive ingredient, early maturity characteristics and the yield of 14 p.eryngii strains were compared. The results showed that the higher yield of these strains were PL 14, PL 17 and PL15; the economic properties of PL 15, PL 16, PL 17 and PL3 strains were better than other strains; the mycelia rate of PL2 and PL 12 mother cultures grew faster than others at 24℃; the higher resistant strains were PL 14, PL7 and PL11; the premature cultivated variety was PL14.
    Esterase isozyme of mycelia of 14 p.eryngii strains were studied by orthogonal slab polyacrylanide gel electrophoresis, the genetic variance and genetic relationship of strains were studied with esterase isozyme techniques and cluster method. The results showed that there was plentiful expression on isozyme band at 0.672 of Rf value. But it was different in the numbers of isozyme bands, position and enzyme activity, individual strains had its peculiar bands. The similarity coefficient of these strains ranged from 0.333 to 1.000. There were various degrees of genetic diversity, however in terms of all the strains tested the degree of genetic variability as a whole was not so plentiful. It indicated that germplasm of p.eryngii was diverse, on the same time, there were no positive connection on one certain degree between the geographical distribution and the genetic variance.
    Cluster analysis based on the coefficient of genetic similarity in the tested strains of p.eryngii was carried out by UPGMA. The results showed that the heterosis was stronger when the cross combinations were produced by most of parent strains in different groups, while the heterosis was weaker when the cross combinations were produced by parent strains in the same group. So parent strains in different groups should be selected in the
    
    
    
    crossbreeding. Ninety-five cross combination were obtained through monospore crossbreeding. When mutually complementary enzyme bands and new enzyme bands of the new cross combinations appeared twenty-two crossbreeding filial generation by comparison of enzyme bands between parent strains and cross combinations, the heterosis of the crossbreeding filial generation was stronger and high quality strains could be select easily.
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