文章摘要
短波紫外线辐照坛紫菜壳孢子制备色彩突变体的新途径
A novel pathway to produce color mutants by short-wavelength ultraviolet ray irradiation on the germinating conchospore in Pyropia haitanensis
投稿时间:2023-01-10  修订日期:2023-02-14
DOI:
中文关键词: 坛紫菜  壳孢子  短波紫外线  色彩突变体  分离
英文关键词: Pyropia haitanensis  conchospores  short-wavelength ultraviolet  color mutants  isolation
基金项目:现代农业产业技术体系建设专项项目(CARS-50)、浙江省农业(水产)新品种选育重大科技专项项目(2021C02069-9)、福建省科技计划项目(2019R1013-8)和温州市科技计划项目( S2020008)共同资助
作者单位邮编
陶伟丽 福建省水产研究所 361000
钟晨辉 福建省水产研究所 
张鹏 浙江省海洋水产养殖研究所 
郭辰涛 福建省水产研究所 
王铁杆 浙江省海洋水产养殖研究所 
林琪* 福建省水产研究所 361000
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中文摘要:
      紫外线诱变是一种快捷有效的紫菜诱变育种方法,但对壳孢子进行的诱变研究鲜有报道。壳孢子萌发时期是坛紫菜减数分裂发生的时期,壳孢子萌发形成的最初四个细胞呈线性排列,形成减数分裂四分体,且发生遗传重组的四分体细胞可以决定叶状体的发育模式和性状分离。因此,诱变产生的色彩突变的嵌合叶状体相比诱变叶状体产生的点状色块,将更易于获得突变细胞。本研究为获得坛紫菜人工色彩嵌合突变体,使用不同剂量(50、100、200、300、400、500和600 J/m2)的短波紫外线(λ=254nm)辐照坛紫菜壳孢子,培养数天后,在壳孢子苗中出现了色彩突变的嵌合叶状体。结果表明,低剂量(50J/m2)的辐射促进壳孢子萌发,而高于100J/m2的辐照剂量,则会抑制壳孢子萌发和分裂。在辐照剂量50~400J/m2范围内,色彩突变体出现的频率随辐照剂量的增加而增加,300J/m2400J/m2剂量辐照时,突变率分别达到15.22%和17.18%。其中,出现色彩突变的嵌合叶状体以2色块嵌合体、3色块嵌合体居多,4色块嵌合体最少,但增加至400J/m2以上时,随辐照剂量的增加,色彩突变体出现的频率反而下降,这表明适宜的诱变剂量为300 J/m2400J/m2。同时短波紫外线(UV-C)辐照也致使色彩突变嵌合体的长宽比下降,采用生物酶解法从色彩嵌合体中分离出了单色的体细胞萌发体。本研究为坛紫菜人工色彩突变体的制备和诱变育种提供了新途径。
英文摘要:
      Ultraviolet mutagenesis is a safe and efficient method to induce mutations in laver, it has the advantages of non-pollution, high efficiency, easy operation, low cost, etc. It has been mainly used for mutagenesis of the filament, protoplast or blade of cultivated Pyropia species, but rarely used in inducing genetic mutants from germinating conchospore. The germination period of conchospores is the period of meiosis in Pyropia haitanensis. The four progeny cells of germinating conchospores are linearly arranged forming a meiotic tetrad. The tetrad cells that undergo genetic recombination can determine the developmental pattern and segregation of traits in the thallus. In this study, short-wave ultraviolet (UV-C) irradiation with different doses (50, 100, 200, 300, 400, 500 and 600 J/m2) was used to induce color mutants during the germination of conchospores in P. haitanensis. The results showed that the low-dose irradiation (50J/m2) promoted the germination of conchospores, while irradiation doses above 100J/m2 inhibited the germination and growth of tetrad germlings. Therefore, low-dose of UV-C irradiation was used in the production to promote the germination of conchospores and improve the production efficiency of laver. In the dose range of 50-400J/m2, the frequency of color chimeras increased with the increase of irradiation intensity. When the dose was 300J/m2 and 400J/m2, and the pigmentation mutation rate was 15.22%, 17.18%, respectively, and the death rate of conchospores was 51.7%, 61%, respectively. In the dose range of 50-500J/m2, with the increase of UV-C irradiation dose, the proportion of 2-color sectored chimera showed a trend of decreasing first and then increasing, and the proportion of 3-color sectored chimera and 4-color sectored chimera showed a trend of increasing first and then decreasing. Among them, the regenerated color chimeras that appear were generally 2-color sectored chimera and 3-color sectored chimera, yet the proportion of 4-color sectored chimera was the least. When the irradiation intensity reached 500J/m2, most of conchospores died, and the death rate was 83.98%, and the frequency of color mutants was significantly lower than that of 300 J/m2 and 400J/m2 dose groups. Those results indicated that the mutagenesis effect was the best when the dose was 300J/m2 or 400J/m2, which was convenient to obtain more abundant genetic recombination and genetic variation in progeny cells. In addition, UV-C irradiation also had a significant effect on the early development of conchospores and phenotypes of pigmentation mutant arranged in tetrad germlings, which was mainly manifested in the large number of color-sectored blades developed from the irradiated conchospores. At the same time, UV-C irradiation retarded the development of cells at the top of conchospore germlings, inhibited the development of cells at the middle base toward the poles, and increased the lateral development, resulting in the decrease in blade aspect ratios. Somatic cell germlings with single colored pigmentation were also isolated from color sectored chimeras by enzymatic hydrolysis. The color species of the five monochromatic mutants obtained were basically consistent with the pigment variant sectors observed on color-sectored thallus, indicating that the obtained color mutants were derived from a single mutant cell on the maternal color-sectored thallus. In conclusion, UV-C irradiations can effectively mutate the conchospores of P. haitanensis, and appropriate irradiation doses (300 J/m2 or 400J/m2) can obtain a certain number of color-sectored thallus. This study provides a novle pathway for preparation of artificial color mutants and mutation breeding in P. haitanensis.
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