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| 坛紫菜诱变及F1代叶状体表型性状分析 |
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张亦弛,王文磊,徐燕,许凯,纪德华,陈昌生,谢潮添
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1.集美大学水产学院 福建 厦门 361021;2.福建省水产生物育种与健康养殖工程研究中心 福建省
发展和改革委员会 福建 厦门 361021;3.农业农村部东海健康养殖重点实验室 福建 厦门 361021;4.农业农村部东海健康养殖重点实验室 福建 厦门 361022;5.农业农村部东海健康养殖重点实验室 福建 厦门 361023;6.农业农村部东海健康养殖重点实验室 福建 厦门 361024;7.农业农村部东海健康养殖重点实验室 福建 厦门 361025;8.农业农村部东海健康养殖重点实验室 福建 厦门 361026;9.农业农村部东海健康养殖重点实验室 福建 厦门 361027
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| 摘要: |
| 突变体是开展坛紫菜(Neoporphyra haitanensis)良种选育和性状遗传调控机理研究的重要材料。为获得坛紫菜突变体，本研究利用不同强度的γ射线辐照(辐射剂量梯度：700、900、1100、1300、1500 Gy)处理野生品系NSD35的幼苗，恢复培养结果显示，γ射线照射导致叶状体部分细胞死亡，且藻体细胞的死亡量随着诱变剂量的增加而升高；同时，突变的细胞数量随着辐射剂量增加呈先增后降的趋势。其中，经1300 Gy处理后藻体的突变细胞最多。之后利用体细胞酶解技术和单克隆技术获得了突变体的纯系藻体，从中初步筛选出性状各异的株系67个，并利用14个表型性状对其中21个株系的F1代进行相关性分析和聚类分析。结果显示，相较于对照组，突变体F1代群体中大部分性状的变异系数增加。突变体的多个性状间存在显著相关性，其中，藻体长度与宽度、鲜重没有显著相关性(P>0.05)；宽度与鲜重、叶形态呈极显著正相关(P<0.01)；鲜重与颜色呈显著负相关(P<0.05)；藻体不同部位的厚度之间存在极显著正相关；叶型与藻体中部、尖端的厚度呈显著负相关。进一步采用系统聚类的方法(遗传距离为20)，将21份材料分成4个主要类群，分别为颜色偏红的藻体组、宽而生物量大的藻体组、薄而日均增长快的藻体组和长而窄的藻体组。综上所述，γ射线对坛紫菜叶状体具有良好的诱变效果，本研究为开展坛紫菜经济性状的遗传调控机理研究以及优良新品种选育提供了基础材料。 |
| 关键词: 坛紫菜 γ射线 酶解 突变体 聚类分析 |
| DOI：10.19663/j.issn2095-9869.20220301001 |
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| Mutagenesis and blade phenotypic traits of Neoporphyra haitanensis F1 lines |
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ZHANG Yichi1,2,3,4, WANG Wenlei1,2,5,6, XU Yan1,2,7,8, XU Kai1,2,9,8, JI Dehua1,2,10,8, CHEN Changsheng1,2,11,8, XIE Chaotian1,2,12,8
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1.Fisheries College, Jimei University, Xiamen, Fujian 361021, China;2.Fujian Engineering Research Center of Aquatic Breeding and Healthy Aquaculture;3.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361021, China;4.Key Laboratory of Healthy Mariculture for the East China Sea,
Ministry of Agriculture and Rural Affairs, Xiamen, Fujian 361021, China;5.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361022, China;6.Key Laboratory of Heal 陠࿗ ꗠ 鞠࿗ ꧰ 飠࿗ 萐࿁ 鉸࿗ 螐࿁ 虰࿗ 衰࿁ 蟘࿗ 襐࿁ 誨࿗ 訰࿁ 谐࿗ 麰࿂ 鳰࿗ ꁐ࿂ 鸰࿗ ꄠ࿂ 齰࿗ 긠࿂ 垰࿖ 릀࿂ 声࿖ 췐࿂ 襀࿗
;7.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361023, China;8.Key Laboratory of Heal Ɛ 蘀䀀ᜀ宋体 es New Roman 虠ဤ ￬ Ɛ 脀䀀ᜀBatang ew Roman 掐ᗡ 襀࿗⏠ᗥ ;9.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361024, China;10.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361025, China;11.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361026, China;12.Fujian Provincial Development and Reform Commission, Xiamen,
Fujian 361027, China
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| Abstract: |
| Neoporphyra haitanensis is a traditional coastal aquaculture species in China; it is rich in essential amino acids, minerals, and vitamins. It is not only beneficial economically, but also adjusts the ecosystem. However, fine varieties of N. haitanensis are lacking even with the enlargement and extension of the N. haitanensis growing area. Since humans began breeding N. haitanensis, most produced strains have been domesticated from wild N. haitanensis collected from rocks. The germplasm collection and usage is considered the bottleneck issue that often causes N. haitanensis quality degradation. Therefore, it is important to breed new varieties. Selective breeding, cross-breeding, and mutation breeding are among the most common methods used to breed new N. haitanensis varieties. Mutation breeding is a quick, simple, and convenient genetic tools. For several decades, natural means to induce genetic diversity have been exploited to breed new varieties, as the naturally occurring mutations are insufficient. One of the most important genetic breakthroughs was the invention of artificial methods to induce mutations. Physical mutagenesis is a safer and more efficient procedure than chemical mutagenesis. To obtain artificial mutants, N. haitanensis (NSD35) gametophytes were treated with different γ-ray irradiation intensities (700 Gy, 900 Gy, 1100 Gy, 1300 Gy, and 1500 Gy). After culture recovery, the results showed that gametophytic cell death increased with the irradiation escalation, with more than 90% cell viability after low radiation dose treatment (700 Gy and 900 Gy). With doses more than 1100 Gy, the cell viability decreased, in which the NSD35 cellular space became larger and a few middle cells died, although no obvious change was observed in tip cells. After induction, the cell morphology changed with higher radiation doses (1300 Gy and 1500 Gy). Some cells exhibited microscopic features, such as enlarged size and alternation from irregular cell shape to circular for some cells. Notably, the cell death rate decreased systematically from the blade base to its tip and from that to its center. Microscopic examination revealed that the number of mutant cells increased and then decreased with the radiation dose rise. The pigmented mutant cells frequency was higher in low radiation doses, further increasing with the rise in irradiation from 700 Gy to 1300 Gy. Nonetheless, the pigmented mutant cells frequency decreased with the highest dose of 1500 Gy. The best mutagenic effect was observed in gametophytes treated with 1300 Gy as they had the largest number of pigmented mutant cells. Cells cultured in vitro were obtained by enzymatic hydrolysis and individual regeneration. Preliminary screening of 67 mutants was performed to construct the mutant library, and 45 pigmented mutant strains, 51 morphological mutants, and 11 other mutants were obtained. Most of the mutants showed two or more mutations of phenotypic character. Twenty-one F1 generation cell lines were screened, and 14 traits (length, width, fresh weight, daily length growth rate, daily width growth rate, daily weight growth rate, length-breadth ratio, base thickness, center thickness, tip thickness, color, base section form, leaf form, saw tooth size, twist, and maturity) were analyzed by correlation analysis and system analysis. The coefficients of variation results showed significant phenotypic differences between F1 and control cells. The F1 coefficients of variation ranged from 8.74% to 59.49%, revealing a moderate variability. Correlation analysis also showed that most of the traits had significant correlations. There was no significant correlation between the length and width or fresh weight, while a significant positive correlation was observed between the width and leaf fresh weight (P<0.01). Significant positive correlations were also found between quantitative traits, while there was no correlation between qualitative and quantitative traits. These allow indirect selection to improve breeding efficiency. Cluster analysis showed that at the Euclidean distance of 20, all 21 individuals analyzed were clustered into four groups. Overall, the phenotypic traits of different groups were significantly different. Mutants are critical materials for studying the genetic regulation of mechanisms involved in the control of economically important N. haitanensis traits. Pigmented mutants are a dominant-marker trait used for breeding that have high value for theoretical research and practical applications. Our study identified many pigmented mutants that can be used to study relevant biological mechanisms. Nevertheless, the variation coefficients of four quantitative characters (length, width, fresh weight, and thickness) of the F1 N. haitanensis generation were less than those of the control group and were biased towards negative variation. For example, more thin leaf mutants were obtained after the gamma rays´ treatment. It is interesting as a thin N. haitanensis is more suitable for automatic processing, and its primary products can be reprocessed with higher added value. In conclusion, after being treated with γ-ray, the N. haitanensis blades were guided to variation enabling the selection and cultivation of new strains, from which strains with improved traits strains were selected. This study provides interesting materials for accelerating the N. haitanensis breeding research and the selection of excellent varieties. |
| Key words: Neoporphyra haitanensis Gamma ray Enzymolysis Mutant Clustering analysis |
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