氮离子注入白金针菇生物学效应的研究
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摘要
离子注入白金针菇及其生物学效应的研究在国内还是首次,具有一定的创新性。本论文的研究目的是研究离子注入对白金针菇的生物学效应的影响,为获得优质、高产的金针菇菌株提供较佳的参数和新的出发菌株,为食用菌育种的学术研究提供重要的参考价值。
在本文中,通过正交优化试验确定出了白金针菇菌丝生长的最佳固体培养基配方,液体发酵试验筛选出了较好的液体发酵配方。在此基础上对白金针菇F3进行了三次N+离子注入。注入结果表明,N+离子注入对白金针菇的生物学效应明显,不同注入剂量的生物学效应不同,呈“马鞍型”曲线变化规律。同时,离子注入会导致白金针菇菌丝细胞发生突变,且突变率高,突变谱宽。通过一系列筛选工作和生物学效应的分析对比,获得了与出发株相比有明显变异特征的正突变株F6、F615和负突变株F62。对他们进行了出菇试验,结果突变菌株F6、F615第一茬菇的平均生物转化率比出发株提高了24.01%,出菇周期缩短了15d,均盖直径小了0.3cm。而突变菌株F62的出菇情况则相反,第一茬菇的平均生物转化率仅为35.38%,比出发菌株下降了39.75%,出菇周期长达97d。最后,总结了一些出菇栽培过程中的经验,为工厂化生产提供一些参考。
The research on biological effects of white flammulina velutipes implantated by nitrogen ion beam was studied for the first time at home and there was some innovations. The intention of this paper was to study the biological effects of white flammulina velutipes implantated by nitrogen ion beam and to offer some important parameters and new strains for the flammulina velutipes with good propoties and high yields. There was also some excellent reference values for the scientifical study of edible mushroom breeding.
In the paper, the best prescription constituents of solid medium to the mycelium growth of white flammulina velutipes was founded by orthogonality optimization experiment and the better prescription constituents of liquid fermentation medium was screened out by liquid fermentation experiment. On this base, the white flammulina velutipes F3 was implanted by nitrogen ion beam three times. The results indicated that there was remarkable biological effects on the white flammulina velutipes implanted by nitrogen ion beam and the effects were different with different doses. The relation between the doses and the survival rate was“saddle modle”curve. At the same time, the mycelial cells was induced by nitrogen ion beam implantation. And the mutation rate was high, the mutation spectrum was wide. The positive mutation strain F6, F615 and the negative mutation strain F62 which had remarkable characters compared with the control strain (F3) were finally screened out through a series of filter work and contrast analysis of biological effects.
The average biotransformation rate for the first fruit of F6, F615 was 24.01% higher than that of the control strain, the fruit cultivation periods was shorter and the diameter of pileus was 0.3 centimeters smaller than that of the control strain. The the mutation strain F62 was the reverse. The average biotransformation rate of F62 for the first fruit was only 35.38% which was 39.75% lower than that of the control strain. The fruit cultivation periods of F62 lasted for 97days. Finally, some experience of the growth and cultivation were sumed up and some referrences for industrialization production were supplied.
在本文中,通过正交优化试验确定出了白金针菇菌丝生长的最佳固体培养基配方,液体发酵试验筛选出了较好的液体发酵配方。在此基础上对白金针菇F3进行了三次N+离子注入。注入结果表明,N+离子注入对白金针菇的生物学效应明显,不同注入剂量的生物学效应不同,呈“马鞍型”曲线变化规律。同时,离子注入会导致白金针菇菌丝细胞发生突变,且突变率高,突变谱宽。通过一系列筛选工作和生物学效应的分析对比,获得了与出发株相比有明显变异特征的正突变株F6、F615和负突变株F62。对他们进行了出菇试验,结果突变菌株F6、F615第一茬菇的平均生物转化率比出发株提高了24.01%,出菇周期缩短了15d,均盖直径小了0.3cm。而突变菌株F62的出菇情况则相反,第一茬菇的平均生物转化率仅为35.38%,比出发菌株下降了39.75%,出菇周期长达97d。最后,总结了一些出菇栽培过程中的经验,为工厂化生产提供一些参考。
The research on biological effects of white flammulina velutipes implantated by nitrogen ion beam was studied for the first time at home and there was some innovations. The intention of this paper was to study the biological effects of white flammulina velutipes implantated by nitrogen ion beam and to offer some important parameters and new strains for the flammulina velutipes with good propoties and high yields. There was also some excellent reference values for the scientifical study of edible mushroom breeding.
In the paper, the best prescription constituents of solid medium to the mycelium growth of white flammulina velutipes was founded by orthogonality optimization experiment and the better prescription constituents of liquid fermentation medium was screened out by liquid fermentation experiment. On this base, the white flammulina velutipes F3 was implanted by nitrogen ion beam three times. The results indicated that there was remarkable biological effects on the white flammulina velutipes implanted by nitrogen ion beam and the effects were different with different doses. The relation between the doses and the survival rate was“saddle modle”curve. At the same time, the mycelial cells was induced by nitrogen ion beam implantation. And the mutation rate was high, the mutation spectrum was wide. The positive mutation strain F6, F615 and the negative mutation strain F62 which had remarkable characters compared with the control strain (F3) were finally screened out through a series of filter work and contrast analysis of biological effects.
The average biotransformation rate for the first fruit of F6, F615 was 24.01% higher than that of the control strain, the fruit cultivation periods was shorter and the diameter of pileus was 0.3 centimeters smaller than that of the control strain. The the mutation strain F62 was the reverse. The average biotransformation rate of F62 for the first fruit was only 35.38% which was 39.75% lower than that of the control strain. The fruit cultivation periods of F62 lasted for 97days. Finally, some experience of the growth and cultivation were sumed up and some referrences for industrialization production were supplied.
引文
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[2] 余增亮等.离子束生物技术引论[M].合肥:安徽科学技术出版社,1998,179.
[3] 吴定,路桂红.低能离子注入法诱变微生物育种[J].中国酿造,2002(增刊),总第123 期:31～33,37
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[5] 周长芳,钦佩.低能重离子生物学研究进展[J].南京林业大学学报(自然科学版), 2003,27(1):81～86.
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[8] 乔长晟,汤凤霞,王文辉.金针菇液体发酵工艺条件的研究[J].宁夏农学院学报,2001,2(22):42～43,82.
[9]Feng,Gong,YaLi,Chen,Ming,Gong etc.Bioresource Technology,Volume 64,Issue 2, May,1998,Pages 153～156.
[10] 于荣利,秦旭升,宋凤菊.金针菇研究概况[J].食用菌学报,2004,11(4):63～68.
[11]Tadjibaeva,Gulnora,Sabirov,Ravshan and Tomita,Toshio.Biochimica et Biophysica Acta (BBA)/Biomembranes,Volume 1467,Issue 2,August 252000,Pages 431～443.
[12] Magae, Yumi and Hayashi, Noriko.FEMS Microbiology Letters,Volume 180,Issue 2,November 15,1999,Pages 331～335.
[13]Sakamoto Y,Ando A,Tamai Y,etal.Protein expressions during fruit body induction of Flammulina velutipes under reduced temperature[J].Mycological Research,2002,106:222～227.
[14] 王德平,王增池.金针菇食品开发现状及其前景[J].食用菌,2000(1),40～41.
[15] 危贵茂,钟卫民,袁诚.金针菇火腿肠的研制[J].肉类工业,2005,(1):16～18.
[16] 冯昆,张东杰,张涧铮等.凝固型金针菇酸奶的研制[J].黑龙江八一农垦大学学报,2005,l7(3):79～8l.
[17] 刘艳芝.我国金针菇遗传育种研究概况[J].长江学报,2003(10):29～41.
[18] 张淑霞.金针菇诱变育种与生料栽培试验[J].食用菌,2003(3):14～15.
[19] 王付转,梁秋霞,李宗伟等.诱变和筛选方法在微生物育种中的应用[J].洛阳师范学院学报,2002(2):95～99.
[20] 李蕤,虞磊,阚劲松.金针菇菌丝断片单细胞诱变育种的研究[J].合肥学院学报(自然科学版),2003,15(1):16～18.
[21] 熊泽,邵伟,李玲等.富硒金针菇菌种的诱变选育[J].三峡大学学报(自然科学版),2004,26(6):574～576.
[22] 杨宗渠,王柏楠,王勤波等.γ射线诱变选育金针菇新菌株[J].核农学报,2002,16(5):325～327.
[23] 金玲,武艳霞,周宗俊等.原生质体辐射诱变培育金针菇新菌株[J].园艺学报,20OO,27(1):65～66.
[24] 李耀维,冯文新,张素梅.He—Ne 激光对金针菇 SOD 高产株的诱变效应[J].激光生物学报,2002,11(4):283～286.
[25] 胡 国 元 . 聚 类 分 析 在 金 针 菇 杂 交 育 种 中 的 应 用 [J]. 微 生 物 学 通报,1998,25(3):130～132.
[26] 胡国元.金针菇杂交育种研究进展[J].湖北民族学院学报(自然科学版),1997,15(3): 27～30.
[27] 彭卫红,肖在勤,甘炳成.金针菇转核育种研究[J].食用菌学报,2001,8(3):1～5.
[28] 彭卫红,肖在勤.白色金针菇新品种金白 1 号的选育[J].西南农业学报,2001,14(增): 75～78.
[29] 孟祥贤,张志光,卿志荣.金针菇担孢子原生质体制备条件的研究[J].生命科学研究[J].2000,4(1):48～52.
[30] 江玉姬,谢宝贵,吴文礼.金针菇的原生质体单核化[J].福建农业大学学报,2001,30 (1):44～47.
[31] 刘维侠,谢宝贵,王秀全等.利用 RAPD-BSA 分子标记技术筛选金针菇色泽基因研究[J]. 中国农学通讯,2005,21(9):54～56.
[32] 高 岛 幸 司 ( 著 ), 黄 年 来 ( 译 ). 高 钙 金 针 菇 的 栽 培 [J]. 中 国 食 用菌,2002,22(2):16～18.
[33] 雷 泓 志 , 张 进 武 , 万 益 琴 等 . 金 针 菇 富 锌 深 层 培 养 的 研 究 [J]. 食 用菌,2001,23(1):11.
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