秦巴山区黑木耳主要栽培种内原生质体分离与融合

doi:10.3969/j.issn.1673-1689.2014.08.011

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秦巴山区黑木耳主要栽培种内原生质体分离与融合
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                        10.3969/j.issn.1673-1689.2014.08.011
                    
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Study on the Separation and Fusion of Protoplasts of Main Auricularia auricula Cultivars in Qinba Mountains

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采用单一变量法研究了黑木耳神农A8与秦单4#原生质体的制备、再生及融合的最佳条件。培养7 d的黑木耳菌丝体用0.5 mol/L甘露醇做稳渗剂，2.0 mol/L的溶壁酶酶解，30 ℃下水浴3.5 h时，黑木耳神农A8和秦单4#原生质体释放量分别达到0.94×107、1.44×107个/mL；选择RM5再生培养基，神农A8和秦单4#原生质体再生率最高，分别为1.91%、1.53%；原生质体融合的最佳条件为：使用30%聚乙二醇（PEG4000）和0.05 mol/L CaCl2溶液（pH 10）为助融剂，35 ℃融合30 min。通过荧光镜检和撷抗反应得到融合子，为黑木耳的遗传育种奠定理论基础。

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Single variable method was utilized to determine the optimum conditions of Auricularia auricula protoplasts preparation, regeneration and fusion in Shenlong A8 and Qindan 4#. After cultured for 7 d, the Auricularia auricula mycelium was treated with 0.5 mol/L mannitol as osmotic stabilizer, digested by 2.0% lywallzyme, then incubated in 30 ℃ water for 3.5 h, and the total amount of released protoplasts reached 0.94×107/mL, 1.44×107/mL in the two varieties, respectively.RM5 regeneration medium was selected and the highest regeneration rates were observed in Shenlong A8 and Qindan 4# protoplasts, which attained 1.91% and 1.53%, respectively.The optimum condition for protoplasts fusion: 30% PEG4000 and 0.05 mol/L CaCl2(pH 10)were added as fusion-aided agent, and incubated in 35 ℃ for 30 min. The resulting fusant was obtained by fluorescence microscopy and antagonistic reaction, which served as the theoretical basis for Auricularia auricular breeding.

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陈文强,王进,邓百万,彭浩,解修超,兰阿峰,刘开辉,丁小维,赵娜娜,钱雪婷,苏晨曦.秦巴山区黑木耳主要栽培种内原生质体分离与融合[J].食品与生物技术学报,2014,33(8):856-864.

CHEN Wenqiang, WANG Jin, DENG Baiwan, PENG Hao, XIE Xiuchao, LAN Afeng, LIU Kaihui, DING Xiaowei, ZHAO Nana, QIAN Xueting, SU Chenxi. Study on the Separation and Fusion of Protoplasts of Main Auricularia auricula Cultivars in Qinba Mountains[J]. Journal of Food Science and Biotechnology,2014,33(8):856-864.

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在线发布日期: 2014-10-19
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