二套小麦-簇毛麦染色体附加系苗期耐盐性及籽粒硒和叶酸的含量

doi:10.13560/j.cnki.biotech.bull.1985.2023-0003

摘要/Abstract

摘要：

簇毛麦（Dasypyrum villosum）作为小麦的近缘种，具有许多优良基因，是改善小麦耐盐性和提高小麦营养品质的理想材料。以小麦-簇毛麦#2和小麦-簇毛麦#3二体附加系为材料，进行耐盐性鉴定及籽粒中硒和叶酸含量的测定。发现小麦-簇毛麦#2附加系和#3附加系在盐胁迫下的萌发率分别为16.67%-43.33%、26.67%-70.00%，均高于对照中国春（6.65%）；盐胁迫下附加系DA3V#3、DA7V#3、DA2V#3和DA5V#2的萌发率分别为70.00%、56.67%、53.33%和43.33%；盐胁迫下附加系的生长正常，株高和根长均大于对照中国春，其中DA4V#2和DA2V#3的株高分别达到15.2和16.1 cm，DA2V#3根长为3.4 cm；在14个附加系和对照中，DA2V#2和DA2V#3籽粒硒含量最高，分别为8.47和7.60 μg/g。小麦-簇毛麦#2附加系和#3附加系籽粒中叶酸含量为9.00-26.10 μg/100 g，大多数附加系高于对照中国春（10.98 μg/100 g），其中，附加系DA4V#2和DA6V#2与对照差异达极显著水平，DA6V#2比对照增加2.4倍。簇毛麦2V#3、3V#3和5V#2染色体可能存在耐盐相关基因，2V染色体可能存在富硒相关基因，6V#2染色体可能携带籽粒叶酸合成的相关基因。

关键词: 小麦-簇毛麦附加系, 耐盐性, 硒含量, 叶酸含量

Abstract:

Dasypyrum villosum as a related species of wheat has many excellent genes, and it is an ideal material to improve the salt tolerance and nutritional quality of wheat. In this study, wheat-D. villosum #2 and wheat-D. villosum #3 disomic addition lines were applied for the identification of salt tolerance and determination of selenium and folic acid contents in seeds. It was found that the germination rates of the two sets of additional lines under salt stress were 16.67%-43.33% and 26.67%-70.00%, respectively, which were higher than the control Chinese Spring（CS）（6.65%）. The germination rates of the addition lines DA3V#3, DA7V#3, DA2V#3 and DA5V#2 under salt stress were 70.00%, 56.67%, 53.33% and 43.33%, respectively. The additional lines grew normally under salt stress, and the plant height and root length of the additional lines under salt stress were higher than those of the control CS, among which the plant height of DA4V#2 and DA2V#3 reached 15.2 cm and 16.1 cm, respectively, and the root length of DA2V#3 was 3.4 cm. The highest grain selenium content was tested in the addition line DA2V#2（8.47 μg/g）and DA2V#3（7.60 μg/g）containing chromosome 2V in the 14 addition lines. The folic acid content in the seeds of wheat-D. villosum additional lines #2 and #3 was 9.00-26.10 μg/100 g, among which most of the addition lines folic acid content were higher than the control（10.98 μg/100 g）. The difference between DA4V#2 or DA6V#2 and the CS was extremely significant in folic acid content, and the folic acid content in DA 6V#2 was 2.4-fold higher than that in CS. The results inferred that there might be salt tolerance related genes on chromosomes 2V#3, 3V#3, and 5V#2, grain selenium enrichment related genes on chromosome 2V, and grain folic acid biosynthesis related genes on chromosome 6V#2.

Key words: wheat-Dasypyrum villosum additional lines, salt tolerance, selenium content, folic acid content

韩志阳, 贾子苗, 梁秋菊, 王轲, 唐华丽, 叶兴国, 张双喜. 二套小麦-簇毛麦染色体附加系苗期耐盐性及籽粒硒和叶酸的含量[J]. 生物技术通报, 2023, 39(8): 185-193.

HAN Zhi-yang, JIA Zi-miao, LIANG Qiu-ju, WANG Ke, TANG Hua-li, YE Xing-guo, ZHANG Shuang-xi. Salt Tolerance at Seedling Stage and Analysis of Selenium and Folic Acid Content in Seeds in Two Sets of Wheat-Dasypyrum villosum Chromosom Additional Lines[J]. Biotechnology Bulletin, 2023, 39(8): 185-193.

图/表 5

表1 小麦-簇毛麦附加系成熟胚在盐胁迫下的萌发情况

Table 1 Germination situation of mature embryos of wheat- Dasypyrum villosum additional lines under salt stress conditions

材料 Materials	接种胚数Total tested mature embryos	发芽株数 Germinated plants	萌发率 Germination rate/%		材料 Materials	接种胚数Total tested mature embryos	发芽株数 Germinated plants	萌发率 Germination rate/%
中国春 DA1V#2 DA2V#2 DA3V#2 DA4V#2 DA5V#2 DA6V#2 DA7V#2	30 30 30 30 30 30 30 30	2 5 8 9 6 13 10 9	6.66 16.67 26.67 30.00 20.00 43.33 33.33 30.00		DA1V#3 DA2V#3 DA3V#3 DA4V#3 DA5V#3 DA6V#3 DA7V#3	30 30 30 30 30 30 30	13 16 21 10 8 14 17	43.33 53.33 70.00 33.33 26.67 46.67 56.67

表2 小麦-簇毛麦附加系在盐胁迫下的生长情况

Table 2 Growth status of mature embryos of wheat- Dasypyrum villosum additional lines under salt stress conditions

材料 Materials	存活株数 Survival plants	平均株高 Average plant height/cm	平均根长 Average root length/cm		材料 Materials	存活株数 Survival plants	平均株高 Average plant height/cm	平均根长 Average root length/cm
中国春 DA1V#2 DA2V#2 DA3V#2 DA4V#2 DA5V#2 DA6V#2 DA7V#2	2 5 8 9 6 13 10 9	9.4±0.26 d 10.9±0.26 c 12.0±0.36 bc 12.8±0.36 b 15.2±0.79 a 15.0±0.20 d 8.1±0.26 a 14.6±1.12 a	2.8±0.20 ab 2.4±0.17 b 2.7±0.26 ab 2.9±0.10 a 3.0±0.20 a 2.9±0.30 a 2.7±0.10 ab 3.0±0.17 a		DA1V#3 DA2V#3 DA3V#3 DA4V#3 DA5V#3 DA6V#3 DA7V#3	13 16 21 10 8 14 17	14.0±0.20 bc 16.1±0.17 a 14.4±0.62 bc 14.1±0.62 bc 9.9±0.10 cd 13.7±0.26 bc 13.4±0.17 c	3.2±0.17 a 3.4±0.10 a 2.8±0.26 bc 2.2±0.26 d 2.7±0.10 c 3.1±0.17 ab 2.8±0.10 bc

图1 小麦-簇毛麦附加系在盐胁迫下的生长状况 a：小麦-簇毛麦附加系1V-7V#2（1：CS；2-8：DA1V#2-DA7V#2）；b：小麦-簇毛麦附加系1V-7V#3（1：CS；2-8：DA1V#3-DA7V#3）。下同

Fig. 1 Growing status of wheat-Dasypyrum villosum additional lines under salt stress conditions a: Wheat-Dasypyrum villosun addition lines 1V-7V#2（1: CS; 2-8: DA1V#2-DA7V#2）. b: Wheat-Dasypyrum villosun addition lines 1V-7V#3（1: CS; 2-8: DA1V#3-DA7V#3）. The same below

图2 小麦-簇毛麦附加系籽粒中硒含量的测定 a：小麦-簇毛麦附加系1V-7V#2；b：小麦-簇毛麦附加系1V-7V#3。*表示在0.05水平上差异显著，**表示在0.01水平上差异显著。下同

Fig. 2 Selenium content analysis in the grains of wheat-Dasypyrum villosun addition lines a: Wheat-Dasypyrum villosun addition lines 1V-7V#2. b: Wheat-Dasypyrum villosun addition lines 1V-7V#3. * indicates significant differences at 0.05 level, and ** indicates significant differences at 0.01 level. The same below

图3 小麦-簇毛麦附加系籽粒中叶酸含量测定结果 a：小麦-簇毛麦附加系1V-7V#2；b：小麦-簇毛麦附加系1V-7V#3

Fig. 3 Folic acid content results in the grains of wheat-Dasypyrum villosun addition lines a: Wheat-Dasypyrum villosun addition lines 1V-7V#2. b: Wheat-Dasypyrum villosun addition lines 1V-7V#3

参考文献 45

[1]	魏益民, 张波, 关二旗, 等. 中国冬小麦品质改良研究进展[J]. 中国农业科学, 2013, 46(20): 4189-4196. doi: 10.3864/j.issn.0578-1752.2013.20.002
	Wei YM, Zhang B, Guan EQ, et al. Advances in study of quality property improvement of winter wheat in China[J]. Sci Agric Sin, 2013, 46(20): 4189-4196.
[2]	檀竹平, 高雪萍. 1997-2016年中国小麦种植区域比较优势及空间分布[J]. 河南农业大学学报, 2018, 52(5): 825-838.
	Tan ZP, Gao XP. Comparative advantage and spatial distribution of wheat in China from 1997 to 2016[J]. J Henan Agric Univ, 2018, 52(5): 825-838.
[3]	Ma MM, Li YC, Xue C, et al. Current situation and key parameters for improving wheat quality in China[J]. Front Plant Sci, 2021, 12: 638525. doi: 10.3389/fpls.2021.638525 URL
[4]	Miranda da Silveira M, Lambrecht Dittgen C, de Souza Batista C, et al. Discrimination of the quality of Brazilian wheat genotypes and their use as whole-grains in human nutrition[J]. Food Chem, 2020, 312: 126074. doi: 10.1016/j.foodchem.2019.126074 URL
[5]	Rhoades J, Loveday J. Salinity in irrigated agriculture[J]. Agronomy, 1990(30): 1089-1142.
[6]	Li HJ, Zhou Y, Xin WL, et al. Wheat breeding in Northern China: achievements and technical advances[J]. Crop J, 2019, 7(6): 718-729. doi: 10.1016/j.cj.2019.09.003
[7]	Hao M, Zhang LQ, Ning SZ, et al. The resurgence of introgression breeding, as exemplified in wheat improvement[J]. Front Plant Sci, 2020, 11: 252. doi: 10.3389/fpls.2020.00252 pmid: 32211007
[8]	Tyrka M, Tyrka D, Wędzony M. Genetic map of Triticale integrating microsatellite, DArT and SNP markers[J]. PLoS One, 2015, 10(12): e0145714. doi: 10.1371/journal.pone.0145714 URL
[9]	Zhang XL, Shen XR, Hao YF, et al. A genetic map of Lophopyrum ponticum chromosome 7E, harboring resistance genes to Fusarium head blight and leaf rust[J]. Theor Appl Genet, 2011, 122(2): 263-270. doi: 10.1007/s00122-010-1441-3 URL
[10]	Cao AZ, Xing LP, Wang XY, et al. Serine/threonine kinase gene Stpk-V, a key member of powdery mildew resistance gene Pm21, confers powdery mildew resistance in wheat[J]. PNAS, 2011, 108(19): 7727-7732. doi: 10.1073/pnas.1016981108 URL
[11]	何中虎, 兰彩霞, 陈新民, 等. 小麦条锈病和白粉病成株抗性研究进展与展望[J]. 中国农业科学, 2011, 44(11): 2193-2215.
	He ZH, Lan CX, Chen XM, et al. Progress and perspective in research of adult-plant resistance to stripe rust and powdery mildew in wheat[J]. Sci Agric Sin, 2011, 44(11): 2193-2215.
[12]	Guo J, Zhang XL, Hou YL, et al. High-density mapping of the major FHB resistance gene Fhb7 derived from Thinopyrum ponticum and its pyramiding with Fhb1 by marker-assisted selection[J]. Theor Appl Genet, 2015, 128(11): 2301-2316. doi: 10.1007/s00122-015-2586-x URL
[13]	Kumar VS, Satish K, Imran S, et al. Transfer of useful variability of high grain iron and zinc from Aegilops kotschyi into wheat through seed irradiation approach[J]. Int J Radiat Biol, 2016, 92(3): 132-139. doi: 10.3109/09553002.2016.1135263 URL
[14]	Zhong GY, Dvorak J. Chromosomal control of the tolerance of gradually and suddenly imposed salt stress in the Lophopyrum elongatum and wheat, Triticum aestivum L. genomes[J]. Theoret Appl Genetics, 1995, 90(2): 229-236. doi: 10.1007/BF00222206 URL
[15]	单雷, 赵双宜, 陈芳, 等. 小麦体细胞杂种山融3号耐盐相关SSR标记的筛选和初步定位[J]. 中国农业科学, 2006, 39(2)225-230
	Shan L, Zhao SY, Chen F, et al. Screening and localization of SSR markers related to salt tolerance of somatic hybrid wheat Shanrong No.3[J]. Sci Agric Sin, 2006, 39(2)225-230
[16]	刘旭, 史娟, 张学勇, 等. 小麦耐盐种质的筛选鉴定和耐盐基因的标记[J]. 2001(9): 948-954.
	Liu X, Shi J, Zhang XY, et al. Screening salt tolerance germplasms and tagging the tolerance gene(s)using microsatellite(SSR)markers in wheat[J]. Chinese Bull Botany, 2001(9): 948-954.
[17]	魏景芳, 秦君, 王淳, 等. 小麦与多枝赖草耐盐纯合易位系的培育及GISH鉴定[J]. 华北农学报, 2004, 19(1): 40-43. doi: 10.3321/j.issn:1000-7091.2004.01.012
	Wei JF, Qin J, Wang C, et al. Development and GISH identification of salt-tolerant translocation lines between wheat and Leymus multicaulis[J]. Acta Agric Boreali Sin, 2004, 19(1): 40-43.
[18]	Ao TGBY, Lang ML, Li YQ, et al. Cloning and expression analysis of cysteine protease gene(MwCP)in Agropyron mongolicum Keng[J]. Genet Mol Res, 2016, 15(1): GMRvol.15.
[19]	Khalil HB, Brunetti SC, Pham UM, et al. Characterization of the caleosin gene family in the Triticeae[J]. BMC Genomics, 2014, 15(1): 239. doi: 10.1186/1471-2164-15-239
[20]	安调过, 许红星, 许云峰. 小麦远缘杂交种质资源创新[J]. 中国生态农业学报, 2011, 19(5): 1011-1019.
	An DG, Xu HX, Xu YF. Enhancement of wheat distant hybridization germplasm[J]. Chin J Eco Agric, 2011, 19(5): 1011-1019. doi: 10.3724/SP.J.1011.2011.01011 URL
[21]	Hanson AD, Gregory JF. Folate biosynthesis, turnover, and transport in plants[J]. Annu Rev Plant Biol, 2011, 62: 105-125. doi: 10.1146/annurev-arplant-042110-103819 pmid: 21275646
[22]	Fox TE, Atherton C, Dainty JR, et al. Absorption of selenium from wheat, garlic, and cod intrinsically labeled with Se-77 and Se-82 stable isotopes[J]. Int J Vitam Nutr Res, 2005, 75(3): 179-186. doi: 10.1024/0300-9831.75.3.179 pmid: 16028633
[23]	Hawkesford MJ, Zhao FJ. Strategies for increasing the selenium content of wheat[J]. J Cereal Sci, 2007, 46(3): 282-292. doi: 10.1016/j.jcs.2007.02.006 URL
[24]	唐玉霞, 王慧敏, 杨军方, 等. 河北省冬小麦硒的含量及其富硒技术研究[J]. 麦类作物学报, 2011, 31(2): 347-351.
	Tang YX, Wang HM, Yang JF, et al. Studies on the selenium content and selenium enriched technique of winter wheat in Hebei Province[J]. J Triticeae Crops, 2011, 31(2): 347-351.
[25]	鲁璐, 季英苗, 李莉蓉, 等. 不同地区、不同品种(系)小麦锌、铁和硒含量分析[J]. 应用与环境生物学报, 2010, 16(5): 646-649.
	Lu L, Ji YM, Li LR, et al. Analysis of Fe, Zn and Se contents in different wheat cultivars(lines)planted in different areas[J]. Chin J Appl Environ Biol, 2010, 16(5): 646-649.
[26]	Riaz B, Liang QJ, Wan X, et al. Folate content analysis of wheat cultivars developed in the North China Plain[J]. Food Chem, 2019, 289: 377-383. doi: S0308-8146(19)30505-9 pmid: 30955626
[27]	Grądzielewska A. The genus Dasypyrum-part 2. Dasypyrum villosum-a wild species used in wheat improvement[J]. Euphytica, 2006, 152(3): 441-454. doi: 10.1007/s10681-006-9245-x URL
[28]	赵万春, 董剑, 陈其皎, 等. 簇毛麦——用于小麦改良的一种野生植物[J]. 草业科学, 2012, 29(10): 1613-1621.
	Zhao WC, Dong J, Chen QJ, et al. Advance in Dasypyrum villosum-a valuable wild species used in wheat improvement[J]. Pratacultural Sci, 2012, 29(10): 1613-1621.
[29]	陈全战, 张边江, 周峰, 等. 簇毛麦染色体对小麦生理指标的影响[J]. 华北农学报, 2010, 25(5): 137-140. doi: 10.7668/hbnxb.2010.05.028
	Chen QZ, Zhang BJ, Zhou F, et al. Effect of Haynaldia villosa chromosome on physiological index of wheat[J]. Acta Agric Boreali Sin, 2010, 25(5): 137-140.
[30]	Xing LP, Di ZC, Yang WW, et al. Overexpression of ERF1-V from Haynaldia villosa can enhance the resistance of wheat to powdery mildew and increase the tolerance to salt and drought stresses[J]. Front Plant Sci, 2017, 8: 1948. doi: 10.3389/fpls.2017.01948 URL
[31]	Zhang RQ, Sun BX, Chen J, et al. Pm55, a developmental-stage and tissue-specific powdery mildew resistance gene introgressed from Dasypyrum villosum into common wheat[J]. Theor Appl Genet, 2016, 129(10): 1975-1984. doi: 10.1007/s00122-016-2753-8 URL
[32]	Qi LL, Pumphrey MO, Friebe B, et al. A novel robertsonian translocation event leads to transfer of a stem rust resistance gene(Sr52)effective against race Ug99 from Dasypyrum[J]. Theor Appl Genet, 2011, 123(1): 159-167. doi: 10.1007/s00122-011-1574-z pmid: 21437597
[33]	Ye XG, Zhang SX, Li SJ, et al. Improvement of three commercial spring wheat varieties for powdery mildew resistance by marker-assisted selection[J]. Crop Prot, 2019, 125: 104889. doi: 10.1016/j.cropro.2019.104889 URL
[34]	Liang XN, Bie XM, Qiu YL, et al. Development of powdery mildew resistant derivatives of wheat variety Fielder for use in genetic transformation[J]. Crop J, 2023, 11(2): 573-583. doi: 10.1016/j.cj.2022.06.012
[35]	罗廷彬, 李彦, 任崴, 等. 新疆盐碱地种植耐盐小麦土壤盐分的变化[J]. 干旱区资源与环境, 2004, 18(S2): 107-113.
	Luo TB, Li Y, Ren W, et al. Studies on soil salinity change by planting salt-tolerant wheat in saline ground in Xinjiang[J]. J Arid Land Resour Environ, 2004, 18(S2): 107-113.
[36]	赵旭, 王林权, 周春菊, 等. 盐胁迫对不同基因型冬小麦发芽和出苗的影响[J]. 干旱地区农业研究, 2005, 23(4): 108-112.
	Zhao X, Wang LQ, Zhou CJ, et al. Effects of salt stress on germination and emergence of different winter wheat genotypes[J]. Agric Res Arid Areas, 2005, 23(4): 108-112.
[37]	彭智, 李龙, 柳玉平, 等. 小麦芽期和苗期耐盐性综合评价[J]. 植物遗传资源学报, 2017, 18(4): 638-645.
	Peng Z, Li L, Liu YP, et al. Evaluation of salinity tolerance in wheat(Triticum aestium)genotypes at germination and seedling stages[J]. J Plant Genet Resour, 2017, 18(4): 638-645.
[38]	王萌萌, 姜奇彦, 胡正, 等. 小麦品种资源耐盐性鉴定[J]. 植物遗传资源学报, 2012, 13(2): 189-194.
	Wang MM, Jiang QY, Hu Z, et al. Evaluation for salt tolerance of wheat cultivars[J]. J Plant Genet Resour, 2012, 13(2): 189-194. doi: 10.13430/j.cnki.jpgr.2012.02.005
[39]	刘慧, 杨月娥, 王朝辉, 等. 中国不同麦区小麦籽粒硒的含量及调控[J]. 中国农业科学, 2016, 49(9): 1715-1728. doi: 10.3864/j.issn.0578-1752.2016.09.008
	Liu H, Yang YE, Wang ZH, et al. Selenium content of wheat grain and its regulation in different wheat production regions of China[J]. Sci Agric Sin, 2016, 49(9): 1715-1728. doi: 10.3864/j.issn.0578-1752.2016.09.008
[40]	Lyons G, Ortiz-Monasterio I, Stangoulis J, et al. Selenium concentration in wheat grain: is there sufficient genotypic variation to use in breeding?[J]. Plant Soil, 2005, 269(1): 369-380. doi: 10.1007/s11104-004-0909-9 URL
[41]	Poblaciones MJ, Santamaría O, García-White T, et al. Selenium biofortification in bread-making wheat under Mediterranean conditions: influence on grain yield and quality parameters[J]. Crop Pasture Sci, 2014, 65(4): 362-369. doi: 10.1071/CP14061 URL
[42]	Liang QJ, Wang K, Shariful I, et al. Folate content and retention in wheat grains and wheat-based foods: effects of storage, processing, and cooking methods[J]. Food Chem, 2020, 333: 127459. doi: 10.1016/j.foodchem.2020.127459 URL
[43]	Liang QJ, Wang K, Liu XN, et al. Improved folate accumulation in genetically modified maize and wheat[J]. J Exp Bot, 2019, 70(5): 1539-1551. doi: 10.1093/jxb/ery453 pmid: 30753561
[44]	Li SJ, Wang J, Wang KY, et al. Development of PCR markers specific to Dasypyrum villosum genome based on transcriptome data and their application in breeding Triticum aestivum-D. villosum#4 alien chromosome lines[J]. BMC Genom, 2019, 20(1): 289. doi: 10.1186/s12864-019-5630-4
[45]	Li SJ, Jia ZM, Wang K, et al. Screening and functional characterization of candidate resistance genes to powdery mildew from Dasypyrum villosum#4 in a wheat line Pm97033[J]. Theor Appl Genet, 2020, 133(11): 3067-3083. doi: 10.1007/s00122-020-03655-4