不同诱变处理对苜蓿叶片细胞显微和超微结构的影响

doi:10.11686/cyxb2017285

草业学报 ›› 2018, Vol. 27 ›› Issue (6): 72-80.DOI: 10.11686/cyxb2017285

不同诱变处理对苜蓿叶片细胞显微和超微结构的影响

冯鹏^1,2, 孙力², 申晓慧^1,2, 李如来², 李增杰², 李志民², 郑海燕², 姜成³, 杨鹤², 刘俊刚², 郭伟², 张英俊^4,*

1.黑龙江省农业科学院博士后工作站,黑龙江哈尔滨150086;
2.黑龙江省农业科学院佳木斯分院,黑龙江佳木斯 154007;
3.佳木斯大学生命科学学院,黑龙江佳木斯 154003;
4.中国农业大学动物科技学院,北京 100193

收稿日期:2017-06-26 修回日期:2017-09-12 出版日期:2018-06-20 发布日期:2018-06-20
通讯作者: * E-mail: zhangyingjun71@hotmail.com
作者简介:冯鹏(1980-),男,内蒙古化德人,副研究员,博士。E-mail:fenggrass@163.com
基金资助:
黑龙江省博士后特别资助项目(LBH-TZ06020)资助

Effect of different mutagenesis methods on microstructure and ultrastructure of alfalfa

FENG Peng^1,2, SUN Li^1,2, SHEN Xiao-hui^1,2, LI Ru-lai², LI Zeng-jie², LI Zhi-min², ZHENG Hai-yan², JIANG Cheng³, YNAG He², LIU Jun-gang², GUO Wei², ZHANG Ying-jun^4,*

1.Postdoctoral Workstation of Heilongjiang Academy of Agricultural Sciences, Harbin 150086, China;
2.Jiamusi Branch of Heilongjiang Academy of Agricultural Sciences, Jiamusi 154007, China;
3.Life Science College of Jiamusi University, Jiamusi University, Jiamusi 154003, China;
4.Animal Science and Technology College of China Agricultural University, Beijing 100193, China

Received:2017-06-26 Revised:2017-09-12 Online:2018-06-20 Published:2018-06-20
Contact: * E-mail: zhangyingjun71@hotmail.com

摘要/Abstract

摘要： 研究⁶⁰Co-γ射线、紫外线、零磁空间和EMS诱变处理下苜蓿SP1代叶片细胞显微、超微结构的变化,筛选最优诱变处理,选育适合高寒地区种植的苜蓿新品系或品种。以公农1号、Wega7F、WL319HQ、敖汉4个苜蓿品种试验材料,设置⁶⁰Co-γ射线150、300、450 Gy射线强度,紫外线30、60、90 min照射时间,甲基磺酸乙酯(EMS) 0.1%、0.2%、0.4% (v/v) 溶液浓度,零磁空间处理180 d;测定苜蓿叶片细胞显微和超微结构。不同诱变处理后4个苜蓿品种叶片厚度均不同程度增加,其中公农1号EMS 0.4% (v/v)浓度下,叶片厚度最大,为446.16 μm,增厚幅度达到25.86%,同时诱变降低了叶脉突起程度;公农1号⁶⁰CO-γ辐射和紫外线处理细胞结构疏松度均高于对照,诱变后Wega7F细胞结构紧密度升高;EMS处理下海绵组织厚度均降低;低剂量诱变处理下[150 Gy ⁶⁰Co-γ,30 min紫外线,0.1% (v/v) EMS],细胞叶绿体出现变形、基粒片层松散、类囊体解体、脂质球增多等现象,高剂量处理[450 Gy ⁶⁰Co-γ,90 min紫外线,0.4% (v/v) EMS]和零磁空间处理下,叶绿体膜会有部分模糊或解体,基粒片层膨胀或模糊、基粒垛叠程度变化随诱变处理种类增加或降低。4种苜蓿品种的叶片厚度诱变后叶片厚度均增加。在一定梯度范围内,⁶⁰CO-γ射线和EMS诱变效率较高,突变体材料丰富,操作简单,适宜诱变育种的实际操作;150 Gy, ⁶⁰Co-γ射线、60 min紫外线、0.4%浓度EMS处理为苜蓿正向变异处理,有利于筛选有益诱变突变体。

关键词: 诱变, 苜蓿, 显微结构, 超微结构, 突变体

Abstract: The overall aim of this research was to identify the optimal mutation method(s) for breeding new alfalfa varieties to cultivate in alpine regions. In this study, we investigated the effects several mutagenesis treatments [⁶⁰COγ-ray irradiation, ultraviolet radiation, ethyl methyl sulfonate (EMS), and magnetic field-free space] on the microstructure and ultrastructure of the leaves of four alfalfa cultivars; Gongnong 1, Wega7F, WL319HQ, and Aohan. The mutagenesis treatments were as follows: ⁶⁰COγ-ray irradiation (150, 300, 450 Gy), ultraviolet radiation (30, 60, 90 min), ethyl methyl sulfonate [0.1%, 0.2%, 0.4% (v/v)], and magnetic field-free space (180 d). The microstructure and ultrastructure of the leaf were analyzed after these treatments. All of the mutagenesis treatments increased leaf thickness. The greatest increase was in Gongnong 1 under the 0.4% EMS (v/v) treatment, where the final leaf thickness was increased by 25.86 μm to 446.16 μm. The degree of leaf vein protuberance was decreased by the mutagenesis treatments. The degree of cell tightness in Gongnong 1 increased after ⁶⁰COγ-ray irradiation and ultraviolet radiation treatments. The cells in the leaves of wega7F were closer together after all the mutagenic treatments. The ultramicrostructure of the chloroplasts in the alfalfa leaves was also observed under different mutagenic treatments. Under lower doses or concentrations of mutagenic treatments [150 Gy ⁶⁰Co-γ, 30 min ultraviolet radiation, 0.1% (v/v) EMS], the chloroplasts were distorted with disrupted membranes, loose grana lamellae, disintegrated thylakoids, and increased numbers of lipid droplets. Under the highest doses or concentrations of mutagenic treatments [450 Gy ⁶⁰Co-γ, 90 min ultraviolet radiation, 0.4% (v/v) EMS, and 180 d magnetic field-free space] the chloroplasts became swollen, distorted, and severely damaged with fractured grana lamellae and disintegrated membrane systems. The numbers of plastids and starch grains in cells were increased under these treatments. Compared with the other treatments, the ⁶⁰COγ-ray irradiation and EMS treatments showed higher mutagenic efficiency, produced more mutant materials, and were simpler to operate. Therefore, we concluded that these were the best mutation methods. The 150 Gy ⁶⁰COγ-ray irradiation, 60 min ultraviolet radiation, and 0.4% EMS treatments were the mutagenic treatments that were most conducive to screening mutants.

Key words: mutagenesis, alfalfa, microstructure, ultrastructure, mutants

冯鹏, 孙力, 申晓慧, 李如来, 李增杰, 李志民, 郑海燕, 姜成, 杨鹤, 刘俊刚, 郭伟, 张英俊. 不同诱变处理对苜蓿叶片细胞显微和超微结构的影响[J]. 草业学报, 2018, 27(6): 72-80.

FENG Peng, SUN Li, SHEN Xiao-hui, LI Ru-lai, LI Zeng-jie, LI Zhi-min, ZHENG Hai-yan, JIANG Cheng, YNAG He, LIU Jun-gang, GUO Wei, ZHANG Ying-jun. Effect of different mutagenesis methods on microstructure and ultrastructure of alfalfa[J]. Acta Prataculturae Sinica, 2018, 27(6): 72-80.

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不同诱变处理对苜蓿叶片细胞显微和超微结构的影响

Effect of different mutagenesis methods on microstructure and ultrastructure of alfalfa

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