NaN_3诱变和盐碱胁迫对苜蓿愈伤组织生长和生理特性的影响
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摘要
为了解NaN_3诱变和盐碱胁迫对苜蓿愈伤组织生长和生理特性的影响,利用叠氮化钠(NaN_3)诱变和盐碱(NaCl、NaHCO_3、Na_2CO_3)胁迫处理"中苜1号"苜蓿愈伤组织,测定相关生长和生理指标。结果表明:随着NaN_3和盐碱浓度的增加,愈伤组织的存活率降低,在NaN_3浓度为3.0 mmol·L~(-1)(处理8 d),愈伤组织存活率为54.55%,此NaN_3浓度和处理时间为NaN_3诱变半致死浓度;在200 mmol·L~(-1)盐碱浓度胁迫下(胁迫12 d),愈伤组织存活率低于5%,此盐碱浓度和胁迫时间为各盐碱致死浓度;除低浓度NaCl胁迫组外,随着NaN_3和盐碱胁迫浓度的增加,愈伤组织生物量降低,NaN_3降低范围为60.85%～81.88%,NaCl胁迫组降低了13.90%～65.63%,NaHCO_3胁迫组降低了21.71%～67.37%,Na_2CO_3胁迫组降低了61.54%～73.20%,盐碱对愈伤组织生物量的影响为NaCl>NaHCO_3>Na_2CO_3。经NaN_3诱变和盐碱胁迫,愈伤组织可溶性蛋白、可溶性糖和脯氨酸含量显著增加,MDA含量和电导率降低,NaN_3、NaN_3+NaCl、NaN_3+Na_2CO_3、NaN_3+NaHCO_3各处理可溶性蛋白含量分别增加了47.07%、53.57%、22.31%、47.69%,可溶性糖含量分别增加了24.79%、162.86%、58.36%、109.54%,脯氨酸含量分别增加了47.78%、162.86%、80.00%、107.25%,MDA含量分别降低了2.35%、11.17%、3.64%、5.54%,相对电导率分别降低了15.13%、17.96%、16.79%、17.28%。NaN_3诱变促进愈伤组织渗透调节物质积累和降低膜的透性。
In order to decipher the effect of NaN_3 mutagenesis and saline-alkali stress on the growth and physiological characteristics of alfalfa callus, we conducted an experiment that the growth and physiological indexes of alfalfa callus, "Zhongmu No.1"were treated by NaN_3 and saline-alkali(NaCl, NaHCO_3, and Na_2CO_3) stresses. The results showed that increasing NaN_3 and salt concentration decreased the survival rate of callus. When the concentration of NaN_3 was at 3 mmol·L~(-1), 8 d treatment decreased the survival rate of callus to 54.55%. This treatment combination is NaN_3 mutagenic semi-lethal conditions. The survival rate of callus was lower than 5% at 200 mmol·L~(-1)of NaN_3 after 12 d treatment that is the lethal concentration of each salt and stress time. In addition to the low concentration of NaCl stress group, the biomass of the callus decreased with the increase of NaN_3 and salt stress concentration, NaN_3 treatment decreased the biomass in a range of 60.85% to 81.88%, the NaCl stress group decreased that by 13.90%～65.63%, the NaHCO_3 stress group decreased that by 21.71%～67.37%, and the Na_2CO_3 stress group decreased that by 61.54%～73.20%. The effect of salt and alkali on the biomass of alfalfa callus ranked in this order: NaCl> NaHCO_3> Na_2CO_3. The content of soluble protein, soluble sugar and proline in the callus increased significantly, and the content of MDA and electrical conductivity decreased by NaN_3 mutation and salt stress. The soluble protein content in treatments of NaN_3, NaN_3+NaCl, NaN_3+ Na_2CO_3 and NaN_3+ NaHCO_3 increased by 47.07%, 53.57%, 22.31%, and 47.69%, respectively. The soluble sugar content increased by 24.79%, 162.86%, 58.36% and 109.54%, respectively.The proline content increased by 47.78%, 162.86%, 80.00%, and 107.25%, respectively. The MDA content decreased by 2.35%, 11.17%, 3.64%, 5.54%, respectively. The relative conductivity decreased by 15.13%, 17.96%, 16.79% and 17.28% in all treatments, respectively. The NaN_3 induced the accumulation of osmotic substances in the callus and decreased the permeability of membrane.
In order to decipher the effect of NaN_3 mutagenesis and saline-alkali stress on the growth and physiological characteristics of alfalfa callus, we conducted an experiment that the growth and physiological indexes of alfalfa callus, "Zhongmu No.1"were treated by NaN_3 and saline-alkali(NaCl, NaHCO_3, and Na_2CO_3) stresses. The results showed that increasing NaN_3 and salt concentration decreased the survival rate of callus. When the concentration of NaN_3 was at 3 mmol·L~(-1), 8 d treatment decreased the survival rate of callus to 54.55%. This treatment combination is NaN_3 mutagenic semi-lethal conditions. The survival rate of callus was lower than 5% at 200 mmol·L~(-1)of NaN_3 after 12 d treatment that is the lethal concentration of each salt and stress time. In addition to the low concentration of NaCl stress group, the biomass of the callus decreased with the increase of NaN_3 and salt stress concentration, NaN_3 treatment decreased the biomass in a range of 60.85% to 81.88%, the NaCl stress group decreased that by 13.90%～65.63%, the NaHCO_3 stress group decreased that by 21.71%～67.37%, and the Na_2CO_3 stress group decreased that by 61.54%～73.20%. The effect of salt and alkali on the biomass of alfalfa callus ranked in this order: NaCl> NaHCO_3> Na_2CO_3. The content of soluble protein, soluble sugar and proline in the callus increased significantly, and the content of MDA and electrical conductivity decreased by NaN_3 mutation and salt stress. The soluble protein content in treatments of NaN_3, NaN_3+NaCl, NaN_3+ Na_2CO_3 and NaN_3+ NaHCO_3 increased by 47.07%, 53.57%, 22.31%, and 47.69%, respectively. The soluble sugar content increased by 24.79%, 162.86%, 58.36% and 109.54%, respectively.The proline content increased by 47.78%, 162.86%, 80.00%, and 107.25%, respectively. The MDA content decreased by 2.35%, 11.17%, 3.64%, 5.54%, respectively. The relative conductivity decreased by 15.13%, 17.96%, 16.79% and 17.28% in all treatments, respectively. The NaN_3 induced the accumulation of osmotic substances in the callus and decreased the permeability of membrane.
引文
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[18] 范小峰,杨颖丽,程转霞.NaCl胁迫下唐古特白刺愈伤组织生理生化变化研究[J].干旱地区农业研究,2009,27(3):203-207.
[19] Amini F,Ghanbarzadeh Z,Askary M.Biochemical and Physiological Response of Salsolaarbuscula,Callus to Salt Stress[J].Iranian Journal of Science & Technology Transactions A Science,2017,41(2):321-328.
[20] 柳福智,羊健麟.盐胁迫对甘草愈伤组织渗透调节的影响[J].植物生理学报,2015,51(7):1038-1044.
[21] Liu T,Staden J V.Growth rate,water relations and ion accumulation of soybean callus lines differing in salinity tolerance under salinity stress and its subsequent relief[J].Plant Growth Regulation,2001,34(3):277-285.
[22] 张玉霞,张立军,王艳树,等.盐碱对芦笋胁迫效应的分析[J].安徽农业科学,2006,34(18):4678-4679.
[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.
[2] 刘倩,高娅妮,柳旭,等.植物对盐碱胁迫的响应机制研究进展[J].生态学报,2017,37(16):1-13.
[3] 张学云,杨丽,范金,等.紫花苜蓿体细胞胚的辐射诱变及耐盐性筛选[J].中国草地学报,2015,37(3):31-36.
[4] 杨静慧,吴志瑜,刘艳军,等.盐胁迫下耐盐苜蓿松散型愈伤组织生理生化特性变化[J].天津农学院学报,2016,23(4):38-42.
[5] 李源,刘贵波,高洪文,等.NaCl胁迫对不同苜蓿种质苗期生长特性的影响[J].华北农学报,2010,25(8):109-116.
[6] Varsha S,Gopal R K.Salinity-induced modulation of growth and antioxidant activity in the callus cultures of miswak (Salvadora persica)[J].Biotech,2013,3(1):11.
[7] Liu L,Fan X D,Wang F W,et al.Coexpression of ScNHX1,and ScVP,in transgenic hybrids improves salt and saline-alkali tolerance in Alfalfa (Medicago sativa L.)[J].Journal of Plant Growth Regulation,2013,32(1):1-8.
[8] 周玉丽,方林森,胡能兵,等.薄荷不定芽诱导及NaN3诱变研究[J].中药材,2016,39(1):1-5.
[9] 付凤玲,李晚忱,荣廷昭,等.用γ射线和叠氮化钠诱变的玉米愈伤组织筛选耐旱和雄性不育材料[J].核农学报,2005,19(5):356-359.
[10] 韩晓玲.小冠花抗L-羟基脯氨酸(Hyp)变异系离体筛选及其耐盐性研究[D].西安:西北大学,2006.
[11] 景建洲.谷子耐盐变异体和草木樨状黄芪抗乙硫氨酸变异体的筛选[D].西安:西北大学,2000.
[12] 王尚军.水杨酸与烯效唑对盐胁迫下黑果枸杞愈伤组织生理生化特性的影响研究[D].兰州:兰州大学,2011.
[13] 王妍,胡胜,付文成,等.一种快速测定可溶性糖的新方法:TBA法[J].井冈山大学学报(自然科学版),2013,(3):37-40.
[14] 冯学金,郭秀娟,杨建春,等.诱变技术在亚麻育种中的应用[J].核农学报,2017,31(7):1310-1316.
[15] Ehsanpour A A,Fatahian N.Effects of salt and proline on Medicago sativa,callus[J].Plant Cell Tissue & Organ Culture,2003,73(1):53-56.
[16] 罗雷,梁燕,崔霞,等.化学诱变剂诱导番茄耐低温突变体的初步研究[J].中国农学通报,2012,28(22):138-143.
[17] 赵邯郸,关淑艳,徐丹丹,等.玉米愈伤组织的耐盐性筛选[J].湖北农业科学,2015,54(1):206-209.
[18] 范小峰,杨颖丽,程转霞.NaCl胁迫下唐古特白刺愈伤组织生理生化变化研究[J].干旱地区农业研究,2009,27(3):203-207.
[19] Amini F,Ghanbarzadeh Z,Askary M.Biochemical and Physiological Response of Salsolaarbuscula,Callus to Salt Stress[J].Iranian Journal of Science & Technology Transactions A Science,2017,41(2):321-328.
[20] 柳福智,羊健麟.盐胁迫对甘草愈伤组织渗透调节的影响[J].植物生理学报,2015,51(7):1038-1044.
[21] Liu T,Staden J V.Growth rate,water relations and ion accumulation of soybean callus lines differing in salinity tolerance under salinity stress and its subsequent relief[J].Plant Growth Regulation,2001,34(3):277-285.
[22] 张玉霞,张立军,王艳树,等.盐碱对芦笋胁迫效应的分析[J].安徽农业科学,2006,34(18):4678-4679.
[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.