LI Hui,XIA Weiwei,HU Ziyu,et al.Identification of Dikaryon-monokaryon Hybrid of Flammulina filiformis by ISSR-SCAR Marker[J].Northern Horticulture,2022,(16):121-127.[doi:10.11937/bfyy.20220445]
利用ISSR-SCAR标记鉴定金针菇双-单杂交子
- Title:
- Identification of Dikaryon-monokaryon Hybrid of Flammulina filiformis by ISSR-SCAR Marker
- 文献标志码:
- A
- 摘要:
- 以2个金针菇菌株为试材，采用原生质体制备与再生技术获得单核体，鉴定其交配型并构建双-单杂交组合，进一步利用ISSR-PCR技术研究亲本、杂交子及其原生质体单核体的遗传差异，以期为金针菇双-单杂交子的准确鉴定提供参考依据。结果表明：2个金针菇菌株分别获得了2种交配型的原生质体单核体，但所占比例差异较大。将FC6的原生质体单核菌株FC6P23与F93配对构建完全亲和双-单杂交组合，获得杂交子FC693。ISSR-PCR扩增图谱结果显示FC693分别含有单核亲本受体核和双核亲本供体核的特异性条带T3和T9，表现为共显性，证明其为真正的杂交子，进一步将特异性条带转化为SCAR标记，利用多重PCR技术可对双-单杂交子进行快速、准确地鉴定。
- Abstract:
- Two strains of Flammulina filiformis were used to create protoplast monokaryons which mating types were determined by mating test.The full compatiable di-mon mating was carried out，and the parents，hybrids and their protoplast monokaryons were used to ISSR-PCR spectrum analysis in order to provide reference for the identification of di-mon hybrid of Flammulina filiformis.The results showed that two types of protoplast monokaryons were obtained for both strains of FC6 and F93 while the ratio of two mating types for monokaryons were different.The hybrid strain of FC693 was obtained by the full compatiable di-mon mating of parent strains of FC6P23 and F93.ISSR spectrum analysis showed that the specific fragment named T3 was detected in both FC693 and FC6P23 while the specific fragment named T9 was detected in both FC693 and F93.These specific fragments were designed by SCAR primers and used to multiplex PCR for hybrid and parent strains.These ISSR-SCAR markers could distinguish the di-mon hybrid accurately and rapidly.
参考文献/References:
[1]WANG P M,LIU X B,DAI Y C,et al.Phylogeny and species delimitation of Flammulina：Taxonomic status of winter mushroom in East Asia and a new European species identified using an integrated approach[J].Mycological Progress,2018,17(9):1013-1030.[2]CHEN J,LI J M,TANG Y J,et al.Genome-wide analysis and prediction of genes involved in the biosynthesis of polysaccharides and bioactive secondary metabolites in high-temperature-tolerant wild Flammulina filiformis[J].BMC Genomics,2021(21):719.[3]刘启燕,戚俊,王卓仁,等.我国金针菇工厂化生产现状与思考[J].中国食用菌,2021,40(12):83-88,92.[4]李天来,许勇,张金霞.我国设施蔬菜、西甜瓜和食用菌产业发展的现状及趋势[J].中国蔬菜,2019(11):6-9.[5]PILOTTI C A,SANDERSON F R,AITKEN E.Sexuality and interactions of monokaryotic and dikaryotic mycelia of Ganoderma boninense[J].Mycological Research,2002,106(11):1315-1322.[6]常堃.双单杂交技术在金针菇工厂化优良栽培菌株选育的应用[D].武汉:华中农业大学,2014.[7]王波,甘炳成,彭卫红,等.金针菇杂交品种:川金3号[J].食用菌,2006(6):15.[8]谭琦,潘迎捷,陈明杰.香菇申香10号菌种的选育与推广[J].食用菌学报,2000,7(3):6-10.[9]王波.金针菇新品种川金33的选育和栽培应用[J].食药用菌,2018,26(1):38-39.[10]刘新锐,杨志超,肖淑霞,等.高产金针菇新品种农金7号[J].园艺学报,2018,45(6):1219-1220.[11]叶明,潘迎捷,陈永萱.香菇完全亲和双单杂交及其先导核的验证[J].南京农业大学学报,1999,22(3):37-40.[12]CHENG S M,LIN F X,XU C.Implication of mating type and cytoplasm factors in asymmetric distribution of two types of monokaryons recovered from dikaryons in Lentinula edodes[J].Journal of Zhejiang University,2008,34(3):237-244.[13]王波,祁丽萍,贾定洪,等.金针菇双-单杂交遗传机理分析[C].北京：第十届全国食用菌学术研讨会,2014.[14]王波,黄忠乾,甘炳成.一种早熟白色金针菇菌株的创制方法[P].中国：CN102177810B，2012-11-14.[15]袁滨,柯丽娜,黄艺宁,等.应用拮抗和ISSR方法综合鉴定灰树花菌株[J].福建农业学报,2019,34(4):393-399.[16]叶明,潘迎捷,陈永萱,等.利用RAPD技术检测香菇双-单杂交后代[J].微生物学通报,2000,27(4):283-286.[17]于浩,葛志豪,徐丽丽,等.双单杂交技术选育长根菇高品质抗病新菌株[J].南方农业学报,2019,50(12):2621-2628.[18]宋莹,张忠伟,刘俊杰,等.ISSR分子标记鉴定香菇杂交菌株试验[J].食用菌,2015(3):16-18.[19]唐利华,肖扬,边银丙.中国黑木耳主要栽培菌株ISSR指纹分析及SCAR标记[J].菌物学报,2008,27(2):243-251.[20]陈雪凤,吴圣进,王灿琴,等.一个鉴别低温刺激型秀珍菇的ISSR-SCAR标记建立与应用[J].热带作物学报,2021,42(2):325-332.[21]王波,祁丽萍,鲜灵.金针菇双单杂交的遗传分析[J].西南农业学报,2012,25(5):1805-1810.[22]王晓敏,吕瑞娜,李长田.应用SRAP、ISSR和TRAP标记构建金针菇分子遗传连锁图谱[J].分子植物育种,2020,18(13):4377-4383.[23]陈鹏,郭成金.金针菇原生质体制备和再生探究[J].江苏农业科学,2014,42(9):200-204.[24]王波,祁丽萍,贾定洪,等.金针菇菌株单核原生质体交配型与菌丝生长分析[J].西南农业学报,2012,25(4):1407-1413.[25]鲍大鹏.担子菌类食用菌交配型位点结构的研究进展[J].菌物学报,2019(12):2061-2077.[26]鲍大鹏.食用菌杂交育种中的科学问题[J].食用菌学报,2020,27(4):1-24.
相似文献/References:
[1]刘善勇,董霞.微磁场对金针菇生长发育的影响[J].北方园艺,2013,37(04):159.
[2]郭欣欣,樊淑华,王永立.金针菇液体培养基的优化研究[J].北方园艺,2014,38(02):146.
GUO Xin-xin,FAN Shu-hua,WANG Yong-li.Study on the Optimization of Liquid Cultural Medium of Flammulina velutipes [J].Northern Horticulture,2014,38(16):146.
[3]连 凯,潘 辉,王迎鑫,等.不同纤维废弃物对金针菇生长发育的影响[J].北方园艺,2014,38(03):138.
LIAN Kai,PAN Hui,WANG Ying-xin,et al.The Influence of Different Lignocellulosic Wastes on the Growth and Development of Flammulina velutipes [J].Northern Horticulture,2014,38(16):138.
[4]李超.金针菇工厂化生产菌糠栽培凤尾菇配方优化研究[J].北方园艺,2013,37(08):157.
LI Chao.Optimization Research on Cultivate Formula of Pleurotus ostreatus Using Cultural Residue in Flammulina velutiper (Fr.) Sing Industrial Production[J].Northern Horticulture,2013,37(16):157.
[5]张腾霄,王 斌,戴 明,等.东北地区规模化生产金针菇的资源利用问题分析[J].北方园艺,2013,37(09):204.
ZHANG Teng-xiao,WANG Bin,DAI Ming,et al.Analysis of the Use of Preponderant Resources and Key Issures on the Realizing Scale Production of Flammulina velutipes in the Northeast of China[J].Northern Horticulture,2013,37(16):204.
[6]林国智.河北北部工厂化金针菇生产的优势性研究[J].北方园艺,2012,36(20):143.
LIN Guo-zhi.Advantage Study on Flammulina Velutipes of Industrialized Production in the North of Hebei Province[J].Northern Horticulture,2012,36(16):143.
[7]王正凤,文生辉.金针菇品种比较试验[J].北方园艺,2014,38(22):150.
WANG Zheng-feng,WEN Sheng-hui.Variety Comparison Experiment of Flammulina velutipes[J].Northern Horticulture,2014,38(16):150.
[8]张城,陈宏伟,朱蕴兰.锌、硒对二种食用菌菌丝体生物量和甘露醇含量的影响[J].北方园艺,2015,39(19):132.[doi:10.11937/bfyy.201519032]
ZHANG Cheng,CHEN Hongwei,ZHU Yunlan.Influence of Zinc and Selenium on Two Kinds of Edible Fungus’ Mycelium Biomass and Mannitol Content[J].Northern Horticulture,2015,39(16):132.[doi:10.11937/bfyy.201519032]
[9]米青荣,刘贵巧,王秋敏,等.邯郸市金针菇品种资源调查及优良品种筛选试验[J].北方园艺,2015,39(20):130.[doi:10.11937/bfyy.201520032]
MI Qingrong,LIU Guiqiao,WANG Qiumin,et al.tudy on Investigation of Flammulina velutipes Variety Resources and Screening of Fine Varieties[J].Northern Horticulture,2015,39(16):130.[doi:10.11937/bfyy.201520032]
[10]梁 梓,张 维 敏,王 容,等.微波辅助法对金针菇水溶性多糖提取工艺的研究[J].北方园艺,2010,34(05):0.[doi:10.11937/bfyy.201005067]
LIANG Zi,ZHANG Wei -min,W ANG Rong,et al.Sdudy on Microwave -assisted Extraction of Soluble Polysaccharide from Flammulina velutipes[J].Northern Horticulture,2010,34(16):0.[doi:10.11937/bfyy.201005067]
备注/Memo
第一作者简介：李慧（1986-），女，硕士，助理研究员，现主要从事食用菌遗传育种等研究工作。E-mail：lihuiviphappy@163.com.责任作者：高春燕（1964-），女，本科，副研究员，现主要从事食用菌育种与栽培技术等研究工作。E-mail：ycsgcy@163.com.基金项目：河北省重点研发计划资助项目（19226368D）；河北省重点研发计划资助项目（21326315D）；河北省农林科学院创新工程资助项目（2022KJCXZX-JZS-9）；国家食用菌产业技术体系资助项目（CARS-24）。收稿日期：2022-02-09