赤芝担孢子交配型因子鉴定及多样性分析
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摘要
以7个栽培赤芝品种为实验材料,以其原生质体单核化单核菌株为标准测试菌株进行交配型因子多样性分析,并采用三轮杂交法、OWE-SOJ技术及核迁移技术对其中赤芝JC和AL菌株的担孢子单核体交配型因子进行准确测定。结果表明,供试的7个栽培赤芝品种中至少存在8种A因子和8种B因子。通过卡方检验发现赤芝AL、韩芝1号、松杉灵芝及Gl.0032+2菌株的单核体分离符合预期,而血芝和无孢灵芝原生质体单核化菌株,以及赤芝JC担孢子的单核菌株存在偏分离现象。采用OWE-SOJ技术与核迁移试验相互印证,可以确定赤芝的4种担孢子单核体的交配型,赤芝JC为:T1(AXBx)、T2(AyBy)、T3(AyBx)和T4(AXBy),赤芝AL为:T1(AXBx)、T2(AyBy)、T3(AXBY)和T4(AyBx)。
The diversity of mating types was analyzed by the protoplast monokaryons of 7 strains of Ganderma lucidum. The mating types of the basidispores of G. lucidum JC and AL were determined by the method of three-cycle mating system, OWE-SOJ technique and nuclear migration test. The results showed that at least 8 A and 8 B factors were found in the 7 test strains. It was found that the monokaryon segregation of AL, Han-zhi No.1, Song-shan and Gl. 0032 +2 was as expected by χ2 test, while the different mating types of protoplasted monokaryon of Xue-zhi and Wu-bao, the basidispores of JC did not display the expected segregation ratio by χ2 test. Four mating types of the basidispores of G. lucidum were determined by OWE-SOJ technique and the nuclear migration test, which verified each other. The monokaryons of JC: T1(AXBx), T2(AyBy), T3(AyBx) and T4(AXBy). The monokaryons of AL: T1(AXBx), T2(AyBy), T3(AXBY) and T4(AyBx).
The diversity of mating types was analyzed by the protoplast monokaryons of 7 strains of Ganderma lucidum. The mating types of the basidispores of G. lucidum JC and AL were determined by the method of three-cycle mating system, OWE-SOJ technique and nuclear migration test. The results showed that at least 8 A and 8 B factors were found in the 7 test strains. It was found that the monokaryon segregation of AL, Han-zhi No.1, Song-shan and Gl. 0032 +2 was as expected by χ2 test, while the different mating types of protoplasted monokaryon of Xue-zhi and Wu-bao, the basidispores of JC did not display the expected segregation ratio by χ2 test. Four mating types of the basidispores of G. lucidum were determined by OWE-SOJ technique and the nuclear migration test, which verified each other. The monokaryons of JC: T1(AXBx), T2(AyBy), T3(AyBx) and T4(AXBy). The monokaryons of AL: T1(AXBx), T2(AyBy), T3(AXBY) and T4(AyBx).
引文
[1]中华人民共和国卫生部药典委员会.中华人民共和国药典[M].北京:中国医药科技出版社,2010:174.
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[13]柯斌榕,吴小平.赤芝与无柄灵芝担孢子的萌发特性比较[J].食药用菌,2015,23(5):282-287.
[14]谢宝贵.倒置显微镜单孢分离法[J].中国食用菌,2008,124(16):56.
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[17]陈莉,李安政,林范学,等.糙皮侧耳担孢子交配型的鉴定[J].微生物学通报,2007,34(6):1 086-1 089.
[18]程水明,林范学.香菇担孢子交配型比例偏分离的遗传分析[J].中国农业科学,2007,40(10):2 296-2 301.
[19]Kawabata H,Magae Y,Sasaki T,et al.Mating type analysis of monokaryons regenerated from protoplast s of Flammulina velutipes[J].Trans Mycol Soc Japan,1992,33(2):243-247.
[20]Liu S R,Ke B R,Zhang W R,et al.Breeding of new Ganoderma lucidum strains simultaneously rich in polysaccharides and triterpenes by mating basidiospore-derived monokaryons of two commercial cultivars[J].Scientia Horticulturae,2017,216:58-65.
[21]吴弘毅.灵芝交配型与出菇相关基因之选殖与分析[D].台北:台湾大学,2005:68-85.
[22]Liu D B,Gong J,Dai W K,et al.The Genome of Ganderma lucidum provide insights into triterpense biosynthesis and wood degradation[J].Plos One,2012,7(5):1-13.
[23]Chen S L,Xu J,Liu C,et al.Genome sequence of the model medicinal mushroom Ganoderma lucidum[J].Nature Comunications,2012,3(2):913-919.
[2]林志彬.灵芝的现代研究[M].3版.北京:北京大学医学出版社,2007:3-8.
[3]李荣正,何云庆.灵芝抗衰老机理与活性成分研究[J].北京医科大学学报,1991,23(6):474.
[4]解跃华,陈跃辉,刘义彬,等.灵芝多糖对实验大鼠高血脂症的预防作用[J].食品研究与开发,2006,27(6):141-142.
[5]陈若芸,于德泉.灵芝三萜化学成分研究进展[J].药学学报,1990,25(12):940-953.
[6]丁一新,赵明文.灵芝lz-8基因的克隆和原核表达及LZ-8蛋白的免疫印迹检测[J].菌物学报,2006,25(4):587-591.
[7]John R.Genetics of sexuality in higher fungi[M].New York:Ronald Press,1966.
[8]Raper J R,Krongelb J R,Baxter G S.The number and distribution of incompatibility factors in Shizophyllum commune[J].The American Naturalist,1958,92:221-232.
[9]Banerjee S,Sarker A.Studies on heterothallism-ⅣGanoderma lucidum(Leyss)Karst.Proc[J].Indian Acad Sci,1958,49:94-98.
[10]陈裕新,夏志兰,刘鹏,等.灵芝群体交配基因型分析[J].中国农学通报,2012,28(10):213-218.
[11]柯斌榕,卢政辉,吴小平,等.我国南方部分灵芝产区栽培品种调查鉴定与分析[J].福建农业学报,2016,31(12):1 329-1 333.
[12]吴小平,刘方,谢玉荣,等.灵芝原生质体单核化杂交育种[J].中国农学通报,2011,25(23):64-69.
[13]柯斌榕,吴小平.赤芝与无柄灵芝担孢子的萌发特性比较[J].食药用菌,2015,23(5):282-287.
[14]谢宝贵.倒置显微镜单孢分离法[J].中国食用菌,2008,124(16):56.
[15]董洪新,蔡德华,李玉.猪肚菇担孢子交配型的分析[J].微生物学报,2010,37(11):1 617-1 620.
[16]Darmono T W.Morphological characteristics of incompatibility reactions and evidence for nuclear migration in armillaria mellea[J].Mycologia,1992,84(3):367-375.
[17]陈莉,李安政,林范学,等.糙皮侧耳担孢子交配型的鉴定[J].微生物学通报,2007,34(6):1 086-1 089.
[18]程水明,林范学.香菇担孢子交配型比例偏分离的遗传分析[J].中国农业科学,2007,40(10):2 296-2 301.
[19]Kawabata H,Magae Y,Sasaki T,et al.Mating type analysis of monokaryons regenerated from protoplast s of Flammulina velutipes[J].Trans Mycol Soc Japan,1992,33(2):243-247.
[20]Liu S R,Ke B R,Zhang W R,et al.Breeding of new Ganoderma lucidum strains simultaneously rich in polysaccharides and triterpenes by mating basidiospore-derived monokaryons of two commercial cultivars[J].Scientia Horticulturae,2017,216:58-65.
[21]吴弘毅.灵芝交配型与出菇相关基因之选殖与分析[D].台北:台湾大学,2005:68-85.
[22]Liu D B,Gong J,Dai W K,et al.The Genome of Ganderma lucidum provide insights into triterpense biosynthesis and wood degradation[J].Plos One,2012,7(5):1-13.
[23]Chen S L,Xu J,Liu C,et al.Genome sequence of the model medicinal mushroom Ganoderma lucidum[J].Nature Comunications,2012,3(2):913-919.