不同温度和水分组合对川黄芪种子萌发和生理活性的影响_张松林.pdf

分子植物育种，2023 年，第 21 卷，第 3 期，第 966-977 页Molecular Plant Breeding,2023,Vol.21,No.3,966-977研究报告Research Report不同温度和水分组合对川黄芪种子萌发和生理活性的影响张松林王晓宇郭俊霞*李青苗*吴萍张美罗冰王洪苏四川省中医药科学院,中药材品质及创新中药研究四川省重点实验室,成都,610041*共同通信作者,;摘要为了探讨不同温度、水分组合对川黄芪种子萌发和生理活性的影响，以川黄芪种子为材料，设置温度(低温 5,常温 15,高温 25)和湿度(低湿 10%,中湿 30%,高湿 60%)复合 9 个双因素处理，随处理时间(7,14,21 d)测定发芽率和 5 项生理指标。结果表明，不同温度和水分组合均促进了川黄芪种子萌发，其中常温/低湿处理第 14 天的萌发率最高，达 70%。在整个试验过程中，可溶性蛋白(SP)含量逐渐降低，并且湿度越高，SP 含量越低；超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)的活性逐渐升高，并且常温组合的抗氧化酶活性高于低温和高温组合；此外，低温组合和高温组合的丙二醛(MDA)含量明显高于常温组合，说明川黄芪种子萌发后受到低温和高温胁迫，常温条件最适宜萌发。主体间交互检验发现，温度、水分、胁迫时间对 SP、SOD、POD、CAT 和 MDA 均呈极显著影响；主成分分析表明，SOD 与 MDA 可作为评价川黄芪种子生理活性的主要指标。综上表明，常温/低湿处理 14 d 是川黄芪萌发的最佳条件，此时川黄芪种子的生理活性最强。研究结果可为提高川黄芪种子的萌发率提供理论基础。关键词川黄芪种子;萌发;温度和水分;生理活性Effect of Different Temperature and Moisture Combinations on Seed Ger-mination and Physiological Activity of ChuanhuangqiZhangSonglinWangXiaoyuGuo Junxia*Li Qingmiao*Wu PingZhangMeiLuo BingWangHongsuSichuan Provincial Key Laboratory of Quality and Innovation Research of Chinese Materia Medica,Sichuan Academy of Chinese Medicine Sciences,Chengdu,610041*Co-corresponding authors,;DOI:10.13271/j.mpb.021.000966AbstractIn order to explore the effects of different temperature and water combinations on the germination andphysiological activities of Chuanhuangqi seeds were treated with nine two-factor treatments of temperature(lowtemperature5,roomtemperature15,hightemperature25)andmoisturecontent(lowmoisture10%,mediummoisture 30%,high moisture 60%),germination percentage and five physiological indicators were measured withthe processing time(7,14,21 d).The results showed that different temperature and moisture combinations pro-moted Chuanhuangqi seeds germination,with the highest germination rate of 70%at the 14th day of the room tem-perature/low humidity treatment.During the experiment,the soluble protein(SP)content gradually decreased,andthe higher the humidity,the lower the SP content;and the activities of superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)gradually increased,and the antioxidant enzyme activities of the room-temperature com-bination were higher than those of the low-temperature combination and the high-temperature combination;inaddition,the MDA contents of the low temperature combination and the high temperature combination were sig-基金项目：本研究由四川省科技计划重点研发项目(2021YFS0043)资助引用格式：Zhang S.L.,Wang X.Y.,Guo J.X.,Li Q.M.,Wu P.,Zhang M.,Luo B.,and Wang H.S.,2023,Effect of different tempera-ture and moisture combinations on seed germination and physiological activity of Chuanhuangqi,Fenzi Zhiwu Yuzhong(MolecularPlant Breeding),21(3):966-977.(张松林,王晓宇,郭俊霞,李青苗,吴萍,张美,罗冰,王洪苏,2023,不同温度和水分组合对川黄芪种子萌发和生理活性的影响,分子植物育种,21(3):966-977.)nificantly higher than the room temperature.In addition,the MDA content of the low-temperature and high-temperaturecombinationswassignificantlyhigherthanthatoftheroom-temperaturecombination,indicatingthatChuan-huangqiseedsweresubjectedtorelativelyhightemperaturestressaftergermination,androom-temperatureconditionsweremostsuitableforgermination.Theinter-subjectinteractiontestrevealedthattemperature,moistureandprocess-ing time had highly significant effects on SP,SOD,POD,CAT and MDA;the principal component analysis showedthat SOD and MDA could be used as the main indicators to evaluate the physiological activities of Chuanhuangqiseeds.In summary,it was shown that 14 d of room temperature/low humidity treatment was the optimum conditionfor the germination of Chuanhuangqi seeds,at which time the physiological activity of Chuanhuangqi seeds wasstrongest.The results can provide a theoretical basis for improving the germination rate of Chuanhuangqi seeds.KeywordsChuanhuangqi seeds;Germination;Temperature and moisture;Physiological activity黄芪是豆科(Leguminosae)黄芪属(Astragalus L.)多年生草本植物蒙古黄芪(A.membranaceus var.mon-gholicus)或膜荚黄芪(A.membranaceus)的干燥根(赵明等,2000,中国野生植物资源,19(6):5-9;杨庆珍等,2014;国家药典委员会,2020)。黄芪根部含黄芪甲苷、毛蕊异黄酮葡萄糖苷(Ibrahim et al.,2013;刘洋等,2017;胡妮娜和张晓娟,2021)，具有补气升阳、生津养血和益卫固表等功效(刘德丽等,2014;姜辉等,2020,安徽中医药大学学报,39(5):93-96)，被广泛应用于肾炎、糖尿病和白血病等疾病的治疗(彭卫华和曲强,2001,中国中西医结合肾病杂志,2(10):614-616)。黄芪主要以种子繁殖，但由于其种皮坚硬(黄正清等,2012)以及内源抑制物质(赵敏和王炎,2001;常晖等,2015)，导致发芽率低、出苗不整齐，严重影响了的规模化生产(郭先龙等,2018)。据统计，在正常温度和湿度条件下约有 80%的黄芪种子不能萌发(中国药材公司,1995,科学出版社,pp.497)。前人采用浸种(程瑶等,2017)、浓硫酸(药用植物试验场中草药组,1975,特产研究,(2):44-46)、沸水(段琦梅,2005)、砂磨(卫士美和王量,2010;梁喜龙等,2012,江苏农业科学,40(4):243-245)等方法处理黄芪种皮，取得了良好的研究结果；也有研究者采用不同的温度(贾文秀等,2011)处理黄芪种子来减弱其内源抑制物质活性，但是不同研究得出的最佳萌发温度存在较大的差异(刘宇,2006)，这可能和水分条件有关。前人研究表明，水分是影响种子萌发的先决条件，种子在吸收足够水分将淀粉转化为小分子物质，可溶性蛋白(SP)和可溶性糖(SS)，为萌发提供养分(童文等,2012)。但水分过高会导致活性氧(ROS)过量积累(Apel andHirt,2004;Gapper and Dolan,2006)，此时植物通过提高抗氧化酶(SOD,POD 和 CAT)的活性来清除 ROS，以维持正常的萌发进程(李合生,2000)。川黄芪(Chuanhuangqi)是多花黄芪(A.floridusBenth)的干燥根，其药用成分和主治功能与 中国药典 中黄芪的描述基本一致，在四川药用历史悠久，常作为黄芪的代用品(何盼等,2013;国家药典委员会,2020)。与黄芪一样，川黄芪也存在着种子萌发率低的问题，但是目前对川黄芪种子萌发的研究较少，特别是温度和水分条件对川黄芪种子萌发和生理活性的研究仍未见报道。因此，本研究测定了不同温度和水分条件下川黄芪种子萌发率和 5 项生理指标，希望解析温度和水分条件对川黄芪种子萌发和生理活性的影响，希望为川黄芪工厂化育苗提供技术参考。1结果与分析1.1不同温度和水分组合对川黄芪种子萌发的影响不同温度和水分组合的川黄芪种子的萌发率在第 14 天达到峰值，之后逐渐降低，