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Effect of different mutagensis on microstructure and ultrastructure of alfalfa

Feng Peng, Shen Xiaohui, Li Rulai, Sun Li, Wu Hongda, Shi Chen, Li Zengjie, Li Zhimin, Zheng Haiyan, Li Daoming and Liu Jungang

This study aims to check the effect of mutagenesis on microstructure and ultrastructure of alfalfa (M. sativa L.) leaf. Four alfalfa cultivars were treated with four kinds of mutagenesis methods, and then the leaf microstructure and ultrastructure of the tested alfalfa were determined respectively. The alfalfa cultivars namely i.e. Gongnong 1, Wega7F, WL319HQ and Aohan were used as the test materials. The mutagenesis methods including 60COγ-ray irradiation (150, 300, 450 Gy), ultraviolet (30, 60, 90 min), EMS (ethyl methane sulfonate 0.1%, 0.2%, 0.4% (v/v)) and magnetic field-free space (180 d) were used for the leaf treatment. The results showed that the leaf thickness was increased under mutagenesis treatment, among them Gongnong 1 alfalfa under EMS with 0.4% (v/v) was the highest, its leaf thickness was as much as 446.16μm with an increase of 25.86μm. Wega7F’s cell become closer under all mutagenic treatments. The ultra microstructure of the chloroplast in the alfalfa leaves was observed under different mutagenic treatments. When making the dose or the concentration of mutagenic treatments lower, the chloroplast of the cell showed distortion, ambiguity in membrane, loose in grana lamella, disintegration in thylakoid, increased in lipid ball. Under the highest dose or concentration mutagenic treatment and magnetic field-free space, the chloroplast of the cell displayed swelling and distortion, fracture in grana lamella and disintegration in membrane system. Membrane systems of the chloroplast were disintegrated and its chloroplast structures were damaged more seriously, both plastids and starch grain were increased. 60COγ-ray irradiation and EMS had higher mutagenic efficiency, their mutant materials were more abundant, their operations were simple, which was considered as being the reasonable mutation methods. Being more conducive for screening mutants, 60COγ-ray irradiation with150Gy, ultraviolet for 60mins and EMS with 0.4% were the better mutagenic treatments

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