叠氮化钠诱变对不同大豆种质低温耐受性的影响

田鑫; 钟程; 李性苑

doi:10.19303/j.issn.1008-0384.2020.07.002

叠氮化钠诱变对不同大豆种质低温耐受性的影响

doi: 10.19303/j.issn.1008-0384.2020.07.002

凯里学院大健康学院，贵州　凯里　556011

基金项目: 贵州省教育厅自然科学研究项目（黔教合KY字[2014]310）

详细信息

作者简介:
田鑫（1983−），男，博士，副教授，主要从事作物遗传育种研究（E-mail：tianxin_china@163.com）

中图分类号: S 565.1
计量
- 文章访问数: 1076
- HTML全文浏览量: 333
- PDF下载量: 30
- 被引次数: 0
出版历程
- 收稿日期: 2019-06-19
- 修回日期: 2020-01-20
- 刊出日期: 2020-07-31

Effect of Sodium Azide-induced Mutagenesis on Low-temperature Tolerance of Soybean Germplasms

College of Comprehensive Health, Kai-li University, Kai-li, Guizhou　556011, China

摘要

摘要: 目的利用不同浓度叠氮化钠对3个种质大豆叶芽进行离体诱变，筛选最佳诱导浓度，并对其进行生理生化分析，鉴定诱变后不同种质丛生芽的低温耐受性。方法以常蔬特大王、台湾292、剑河大豆叶芽为材料，采用不同质量浓度叠氮化钠（0.4、0.8、1.0 mmol·L⁻¹）进行离体诱导试验，筛选最佳诱导处理；进一步对经最佳诱导处理（0.8 mmol·L⁻¹、48 h）的叶芽进行4℃低温胁迫处理4 d，在低温胁迫前及胁迫期间共5 d中，每天测定其生理生化指标。结果 0.8 mmol·L⁻¹叠氮化钠处理48 h的死亡率略高于半致死率，为最佳诱导处理。3个种质诱变植株在低温胁迫下的渗透调节物质、光合色素含量均高于对照，种质差异表现为台湾292＞常蔬特大王＞剑河大豆。常温条件下叠氮化钠诱变能显著提高台湾292、剑河大豆的SOD、CAT活性。3个种质的正常植株在低温胁迫下的抗氧化能力存在差异，剑河大豆（SOD、POD活性提升）＞常蔬特大王（POD活性提升）＞台湾292（SOD、POD活性略有提升，CAT活性下降）。叠氮化钠诱变对3个种质的低温抗氧化能力提升水平不一样，与对照植株相比，台湾292（POD活性提升，SOD活性略有下降）＞常蔬特大王（无显著变化）＞剑河大豆（SOD、POD活性下降）。常蔬特大王和台湾292的MDA含量表现为先升高后降低，而剑河大豆一直保持升高趋势，但相对于0 d的MDA含量提高的种质间差异表现为：台湾292＞常蔬特大王＞剑河大豆。结论叠氮化钠诱变可提高3个大豆种质的渗透调节物质、光合色素含量，其中：改良种质常蔬特大王和台湾292提升较多，地方种质剑河大豆提升较少。改良种质台湾292诱变前后及低温处理前后表现较好，抗氧化能力有所提升；常蔬特大王诱变前后及低温处理前后均表现稳定，诱变对其性状改良潜力较小；诱变对未经改良的地方种质剑河大豆抗氧化酶活性改变较大，在常温下为有利变异，在低温下为不利变异。综合以上结论可知，叠氮化钠诱变对不同大豆种质的低温耐受性提升水平存在差异，表现为：台湾292＞常蔬特大王＞剑河大豆。
- 大豆 /
- 叶芽 /
- 低温处理 /
- 叠氮化钠 /
- 诱变
Abstract: Objective By immersing buds in varied concentrations of sodium azide solutions to induce mutagenesis in vitro on 3 soybean germplasms, optimized induction conditions were determined based on the effect on the low-temp tolerance of the mutants under stress. Methods Young buds of Changshutedawang (CSTDW), Taiwan 292, and Jianhe soybean germplasms were immersed in sodium azide solutions of different concentrations for the in vitro mutagenesis induction. After the optimized induction treatment (immersing buds at 0.8mmol sodium azide/L for 48h), the plants were subjected to low-temp stress at 4 ℃ for 4d prior to the physio-biochemical determinations. Results The mortality rate of the treated plants was only slightly higher than the median lethal dose（LD₅₀）. Under the low-temp stress, the contents of osmosis regulating substance and photosynthetic pigment in the mutant plants were higher than those in control. At room temperature, the increases on the germplasm was in the order of Taiwan 292＞CSTDW＞Jianhe, and the SOD and CAT in Taiwan 292 and Jianhe were significantly increased. Under low temperature, the antioxidant capacities differed among the germplasms. Overall, Jianhe was higher than CSTDW and followed by Taiwan 292. Specifically, SOD and POD activity increased in Jianhe, POD increased in CSTDW, and SOD and POD slightly increased and CAT decreased in Taiwan 292. Compared to control, Taiwan 292 had the greatest antioxidant activities showing an increased POD and slightly decreased SOD activity, CSTDW was next in line with no significant change, and Jianhe had the least with decreased activities on both SOD and POD. The MDA contents of CSTDW and Taiwan 292 increased initially followed by a decline, while Jianhe maintained an increasing trend. The overall increased levels ranked Taiwan 292＞ CSTDW＞ Jianhe. Conclusion The induced mutagenesis increased the osmosis regulating substance and photosynthetic pigment contents in the 3 soybean germplasms. The improvement on low-temp tolerance was higher for CSTDW and Taiwan 292 but lower for the local Jianhe soybean. After mutagenesis and low-temp treatment, Taiwan 292 performed well with a heightened antioxidant capacity. On the other hand, Jianhe changed significantly on the enzymatic activity, the condition was favorable for the plants at room temperature but disadvantageous at low temperatures. In all, the low-temp tolerance of the soybean germplasms generated by sodium azide-induced mutagenesis were found to be Taiwan 292＞CSTDW＞Jianhe.
- Soybean /
- buds /
- low-temp stress /
- sodium azide /
- mutagenesis

HTML全文

图 1 3个大豆种质诱变植株与对照植株在低温胁迫下的生理指标CSTDW：常蔬特大王，TW292：台湾292，JHS：剑河大豆。

注：处理间连线上标记*、**分别表示差异显著（P＜0.05）、极显著（P＜0.01）。

Figure 1. Physiological indices on mutant and control soybean germplasms under low-temp stress

Note: * and ** represent significant difference (P＜0.05) and extremely significant difference (P＜0.01), respectively.

下载: 全尺寸图片幻灯片

图 2 3个大豆种质诱变与对照（CK）植株在4℃低温胁迫下的SOD活性

注：同一时间点不同处理间标记不同小写、大写字母分别表示差异显著（P＜0.05）、极显著（P＜0.01）；诱变处理不同时间点标记*、**分别表示各时间点与第0 d差异显著（P＜0.05）、极显著（P＜0.01）；CK不同时间点标记*、**分别表示各时间点与第0 d差异显著（P＜0.05）、极显著（P＜0.01），图2～5下同。

Figure 2. Differences on SOD activity between mutant and control soybean germplasms under 4 ℃ low-temp stress

Note: At same sampling time, different lowercase letters and capitalized letters indicate significant differences (P＜0.05) and extremely significant differences (P＜0.01), respectively, on different treatments. For mutation treatment, * and ** represent significant differences (P＜0.05) and extremely significant differences (P＜0.01), respectively, between sampling time and 0d. In CK, * and ** represent significant differences (P＜0.05) and extremely significant differences (P＜0.01), respectively, between sampling time and 0d. Same for the following.

下载: 全尺寸图片幻灯片

图 3 3个大豆种质诱变植株与对照植株在低温胁迫下的POD活性

Figure 3. POD activities of mutant and control soybean germplasms under low-temp stress

下载: 全尺寸图片幻灯片

图 4 3个大豆种质诱变植株与对照植株在低温胁迫下的CAT活性

Figure 4. CAT activities of mutant and control soybean germplasms under low-temp stress

下载: 全尺寸图片幻灯片

图 5 3个大豆种质诱变植株与对照植株在低温胁迫下的MDA含量

Figure 5. MDA contents of mutant and control soybean germplasms under low-temp stress

下载: 全尺寸图片幻灯片

表 1 叠氮化钠处理对大豆芽诱变的影响

Table 1. Effect of immersing buds in sodium azide solutions on induced mutagenesis of soybean germplasms

品种名称 Cultivar name	浓度Concentration/ （mmol·L⁻¹）	处理时间 time/h	数量 number/ 盒（芽数）	污染数 number of contamination/芽	污染率 rate of contamination/%	死亡数 death number/芽	褐化数browning number/芽	死亡率 mortality rate/%	丛生芽数number of buds
常蔬特王Changshutedawang	0.0	48	30（260）	35	13.5	0	0	0.00	230
	0.4	48	30（256）	26	10.2	70	0	27.36	161
	0.8	48	30（231）	35	15.2	119	0	51.30	80
	1.0	48	30（224）	15	6.7	204	20	100.00	0
台湾 292 Taiwan 292	0.0	48	30（190）	15	7.9	0	0	0.00	197
	0.4	48	30（207）	26	12.6	81	3	39.71	165
	0.8	48	30（219）	30	13.8	158	0	72.10	73
	1.0	48	30（197）	22	11.2	176	21	100.00	0
剑河大豆 Jianhe soybean	0.0	48	30（267）	41	15.4	0	0	0.00	247
	0.4	48	30（244）	30	12.3	83	2	34.24	205
	0.8	48	30（275）	14	5.1	181	10	68.27	131
	1.0	48	30（220）	18	8.2	203	17	100.00	0

下载: 导出CSV

参考文献(29)

[1]	HARTMAN G L, WEST E D, HERMAN T K. Crops that feed the World 2. Soybean: worldwide production, use, and constraints caused by pathogens and pests [J]. Food Security, 2011, 3(1): 5−17. doi: 10.1007/s12571-010-0108-x
[2]	张德荣, 张学君, 孟祥盟, 等. 大豆低温冷害敏感时期试验研究报告 [J]. 吉林农业科学, 1987(1):37−39. ZHANG D R, ZHANG X J, MENG X M, et al. Primary study on the key time for soybean plants to be injured by the lower temperature [J]. Journal of Jilin Agricultural Sciences, 1987(1): 37−39.(in Chinese)
[3]	张德荣, 张学君. 大豆低温冷害试验研究报告 [J]. 大豆科学, 1988, 7(2):125−132. ZHANG D R, ZHANG X J. Study on cool injury of soybean [J]. Soybean Sciences, 1988, 7(2): 125−132.(in Chinese)
[4]	GAYNOR L G, LAWN R J, JAMES A T. Agronomic studies on irrigated soybean in southern New South Wales. I. Phenological adaptation of genotypes to sowing date [J]. Crop & Pasture Science, 2011, 62(12): 1056−1066.
[5]	YAMAGUCHI N, YAMAZAKI H, OHNISHI S, et al. Method for selection of soybeans tolerant to seed cracking under chilling temperatures [J]. Breeding Science, 2014, 64(1): 103−108. doi: 10.1270/jsbbs.64.103
[6]	张东辉, 杨青春, 耿臻, 等. 我国大豆育种的现状分析 [J]. 农村经济与科技, 2017, 28(14):36. doi: 10.3969/j.issn.1007-7103.2017.14.028 ZHANG D H, YANG Q C, GENG Z, et al. Current situation of soybean breeding in China [J]. Rural Economy and Science-Technology, 2017, 28(14): 36.(in Chinese) doi: 10.3969/j.issn.1007-7103.2017.14.028
[7]	钮力亚, 于亮, 付晶, 等. 叠氮化钠在农作物育种中的应用 [J]. 河北农业科学, 2010, 14(12):52−53, 57. doi: 10.3969/j.issn.1088-1631.2010.12.020 NIU L Y, YU L, FU J, et al. Application of sodium azide in crops breeding [J]. Journal of Hebei Agricultural Sciences, 2010, 14(12): 52−53, 57.(in Chinese) doi: 10.3969/j.issn.1088-1631.2010.12.020
[8]	苑平, 吴娟娟, 李先信, 等. 叠氮化钠对纽荷尔脐橙腋芽的半致死浓度、生理影响和诱变效率研究 [J]. 湖南师范大学自然科学学报, 2018, 41(1):30−35. YUAN P, WU J J, LI X X, et al. Study on semi-lethal concentration, physiological effects and mutagenic efficiency of sodium azide on axillary buds in newhall navel orange(Citrus sinensis osbeck) [J]. Journal of Natural Science of Hunan Normal University, 2018, 41(1): 30−35.(in Chinese)
[9]	SUGIHARA N, HIGASHIGAWA T, ARAMOTO D, et al. Haploid plants carrying a sodium azide-induced mutation (fdr1) produce fertile pollen grains due to first Division restitution (FDR) in maize (Zea mays L.) [J]. Theoretical and Applied Genetics, 2013, 126(12): 2931−2941. doi: 10.1007/s00122-013-2183-9
[10]	姜振峰, 刘志华, 李文滨, 等. 叠氮化钠对大豆M₁的生物学诱变效应 [J]. 核农学报, 2006, 20(3):208−210. doi: 10.3969/j.issn.1000-8551.2006.03.011 JIANG Z F, LIU Z H, LI W B, et al. M₁ mutagenic effect on soybean induced by NaN₃ [J]. Journal of Nuclear Agricultural Sciences, 2006, 20(3): 208−210.(in Chinese) doi: 10.3969/j.issn.1000-8551.2006.03.011
[11]	李明飞, 谢彦周, 刘录祥, 等. 叠氮化钠诱变普通小麦陕农33突变体库的构建和初步评估 [J]. 麦类作物学报, 2015, 35(1):22−29. doi: 10.7606/j.issn.1009-1041.2015.01.04 LI M F, XIE Y Z, LIU L X, et al. Construction and preliminary assessment of a mutant library of common wheat cultivar shaannong 33 mutated with sodium azide [J]. Journal of Triticeae Crops, 2015, 35(1): 22−29.(in Chinese) doi: 10.7606/j.issn.1009-1041.2015.01.04
[12]	孔佑涵, 苑平, 吴娟娟, 等. 叠氮化钠处理纽荷尔脐橙腋芽的诱变效应研究 [J]. 分子植物育种, 2016, 14(12):3489−3495. KONG Y H, YUAN P, WU J J, et al. Study on the mutagen effect of NaN₃ in axillary bud of newhall navel orange (Citrus sinensis osbeck) [J]. Molecular Plant Breeding, 2016, 14(12): 3489−3495.(in Chinese)
[13]	韩伟, 魏岳荣, 盛鸥, 等. ‘巴西蕉’离体芽的化学诱变和抗镰刀菌酸材料的筛选 [J]. 核农学报, 2012, 26(9):1237−1243. HAN W, WEI Y R, SHENG O, et al. Chemical mutation and screening for tolerance to fusaric acid on shoot tip of Musa AAA Cavendish cv. ‘baxijiao’ [J]. Acta Agriculturae Nucleatae Sinica, 2012, 26(9): 1237−1243.(in Chinese)
[14]	李波, 贾秀峰, 高美玲, 等. 诱变苜蓿愈伤组织抗寒性研究 [J]. 草地学报, 2004, 12(2):95−97. doi: 10.11733/j.issn.1007-0435.2004.02.004 LI B, JIA X F, GAO M L, et al. A research on the cold-resistance of induced alfalfa calluses [J]. Acta Agrestia Sinica, 2004, 12(2): 95−97.(in Chinese) doi: 10.11733/j.issn.1007-0435.2004.02.004
[15]	李波, 白庆武, 马兰, 等. 苜蓿抗性变异细胞系的筛选 [J]. 草业科学, 2003, 20(4):5−9. LI B, BAI Q W, MA L, et al. The selection of alfalfa resistance variation cells [J]. Pratacultural Science, 2003, 20(4): 5−9.(in Chinese)
[16]	钱玉源, 韩轩, 刘祎, 等. 叠氮化钠(NaN₃)诱变在作物性状改良中的应用 [J]. 安徽农业科学, 2017, 45(35):136−138, 141. doi: 10.3969/j.issn.0517-6611.2017.35.041 QIAN Y Y, HAN X, LIU Y, et al. Application of sodium azide (NaN₃) mutation in crop character improvement [J]. Journal of Anhui Agricultural Sciences, 2017, 45(35): 136−138, 141.(in Chinese) doi: 10.3969/j.issn.0517-6611.2017.35.041
[17]	李合生. 植物生理生化实验原理和技术[M]. 北京: 高等教育出版社, 2000.
[18]	高兰英, 马庆. 诱发突变技术在小麦育种研究中的应用 [J]. 山西农业科学, 2009, 37(6):7−12. doi: 10.3969/j.issn.1002-2481.2009.06.002 GAO L Y, MA Q. Mutants induction technology and its application in wheat breeding improvement [J]. Journal of Shanxi Agricultural Sciences, 2009, 37(6): 7−12.(in Chinese) doi: 10.3969/j.issn.1002-2481.2009.06.002
[19]	徐冠仁. 植物诱变育种学 [M]. 北京: 中国农业出版社, 1996.
[20]	BHAGWAT B, DUNCAN E. Mutation breeding of banana cv. Highgate (Musa spp., AAA Group) for tolerance to Fusarium oxysporum f. sp. cubense using chemical mutagens [J]. Scientia Horticulturae, 1998, 73(1): 11−22. doi: 10.1016/S0304-4238(97)00141-6
[21]	喻晓. 大百合无性系突变体诱导及其鉴定研究 [D]. 雅安: 四川农业大学, 2008. YU X. The study on induction and identification of mutations in cloned line of Cardiocinum giganteum [D]. Yaan, China: Sichuan Agricultural University, 2008. (in Chinese)
[22]	马玉涵, 赵岩, 张强, 等. 叠氮化钠诱变对离体蝴蝶兰类原球茎生理的影响 [J]. 核农学报, 2010, 24(2):411−414, 301. doi: 10.11869/hnxb.2010.02.0411 MA Y H, ZHAO Y, ZHANG Q, et al. Effect of azide sodium in mutagenesis on physiological traits of Phalaenosis protocorm-like body in vitro [J]. Journal of Nuclear Agricul Turae Sciences, 2010, 24(2): 411−414, 301.(in Chinese) doi: 10.11869/hnxb.2010.02.0411
[23]	温日宇, 刘建霞, 宋亚静, 等. 叠氮化钠对绿豆种子和幼苗生长的诱变效应 [J]. 山西农业科学, 2017, 45(12):1933−1936. doi: 10.3969/j.issn.1002-2481.2017.12.09 WEN R Y, LIU J X, SONG Y J, et al. Mutagenic effects of sodium azide on the growth of mung bean seeds and seedlings [J]. Journal of Shanxi Agricultural Sciences, 2017, 45(12): 1933−1936.(in Chinese) doi: 10.3969/j.issn.1002-2481.2017.12.09
[24]	张兰. 苹果砧木组培苗耐盐诱变及筛选技术研究 [D]. 保定: 河北农业大学, 2002. ZHANG L. Studies on screening techniques for salt tolerance evaluation of induced mutants of apple stock in vitro [D]. Baoding, China: Hebei Agricultural University, 2002. (in Chinese)
[25]	吕伯钦, 曾昭慧, 邓海, 等. 叠氮化钠的毒性研究 [J]. 卫生研究, 1992, 21(5):228−231, 278-279. LV B Q, ZENG Z H, DENG H, et al. Studies on toxicity of sodium azide [J]. Journal of Hygiene Research, 1992, 21(5): 228−231, 278-279.(in Chinese)
[26]	KIM T, JUNG Y, NA B, et al. Molecular cloning and expression of Cu/Zn-containing superoxide dismutase from Fasciola hepatica [J]. Infection and Immunity, 2000, 68(7): 3941−3948. doi: 10.1128/IAI.68.7.3941-3948.2000
[27]	李波, 邬婷婷. NaN₃诱变和盐碱胁迫对苜蓿愈伤组织生长和生理特性的影响 [J]. 干旱地区农业研究, 2019, 37(2):130−135, 143. doi: 10.7606/j.issn.1000-7601.2019.02.19 LI B, WU T T. Effects of NaN₃ mutagenesis and saline-alkali stress on growth and physiological characteristics of alfalfa callus [J]. Agricultural Research in the Arid Areas, 2019, 37(2): 130−135, 143.(in Chinese) doi: 10.7606/j.issn.1000-7601.2019.02.19
[28]	刘建霞, 侍亚敏, 温日宇, 等. 晋藜1号种子及幼苗对叠氮化钠诱变的响应 [J]. 种子, 2018, 37(1):80−83. LIU J X, SHI Y M, WEN R Y, et al. Response of sodium azide mutagenesis on seeds and seedlings of Jinli No.1 quinoa [J]. Seed, 2018, 37(1): 80−83.(in Chinese)
[29]	宇克莉, 邹婧, 邹金华. 镉胁迫对玉米幼苗抗氧化酶系统及矿质元素吸收的影响 [J]. 农业环境科学学报, 2010, 29(6):1050−1056. YU K L, ZOU J, ZOU J H. Effects of cadmium stress on antioxidant enzyme system and absorption of mineral elements in maize seedlings [J]. Journal of Agro-Environment Science, 2010, 29(6): 1050−1056.(in Chinese)