NaN_3和紫外线复合诱变对苜蓿愈伤组织在干旱和盐胁迫下生长的影响
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摘要
以图牧2号苜蓿愈伤组织为材料,对其进行不同浓度NaN_3及紫外线诱变剂量和不同浓度NaCl及PEG胁迫处理,观察愈伤组织生长状况,以愈伤组织的存活率确定各处理的半致死浓度(LC_(50))或剂量(LD_(50))和致死浓度(LC_(100))或剂量(LD100),并对愈伤组织进行脯氨酸、可溶性糖、丙二醛含量和CAT酶活性的测定。结果表明:诱变和胁迫影响了苜蓿愈伤组织生长,随NaN_3浓度和紫外线诱变剂量的增加,愈伤组织的存活率下降,NaN_3的LC_(50)和LC_(100)分别为4.0×10~(-3)mol·L~(-1)和5.5×10~(-3)mol·L~(-1),在距离为20cm时,紫外线诱变LD_(50)和LD_(100)分别为6、8min;随NaCl和PEG选择压力的加大,愈伤组织的存活率下降,NaCl的LC_(50)和LC_(100)分别为1.5%和2.0%,PEG的LC_(50)和LC_(100)分别为25%和30%。各诱变和胁迫处理比较,经NaN3和紫外线复合诱变及PEG和NaCl双重选择压力处理愈伤组织存活率最低为16%。NaN3和紫外线复合诱变的愈伤组织脯氨酸、可溶性糖含量、CAT酶活性增加,丙二醛含量下降。诱变和逆境胁迫筛选的苜蓿愈伤组织,可以提高苜蓿愈伤组织对盐旱的抵抗能力,可为利用体细胞扩大苜蓿抗逆性变异来源提供基础。
Alfalfa callus of Tumu No.2 was used as experimental material,callus was mutated with different concentrations of NaN_3 and different dose of UV,and treated with Na Cl and PEG stress at different concentrations. The growth status of the callus was observed. The semi lethal( LC_(50)) and lethal( LC_(100)) concentration( dose) of each treatment was determined according to the survival rate of callus. The content of proline,soluble sugar,MDA and the activity of CAT in callus were assayed. The results showed that the growth of alfalfa callus was affected by mutation and stress.The survival rate of callus had declined with the increase of NaN_3 concentration and UV irradiation dose.The semi lethal and lethal concentrations of NaN_3 was 4.0 and 5.5×10~(-3) mol·L~(-1) respectively. At the distance of 20 cm,the semi lethal and lethal doses of UV induced mutation was 6 and 8 min. The survival rate of callus declined with the concentration of Na Cl and PEG increasing. The lethal and lethal concentrations of Na Cl was 1.5% and 2% respectively. The lethal and lethal concentrations of PEG was 25% and 30% respectively. Compared with different mutation and stress treatments,the survival rate of callus was 16% treated by NaN_3 and UV combined mutagenesis and PEG and Na Cl double stress treatment. The content of proline,soluble sugar,CAT enzyme increased and MDA content decreased in the callus under NaN_3 and UV combined mutagenesis. Screening of alfalfa callus by mutagenesis and stress could improve the salt and drought resistance. It provides the basis for using somatic cells to expand the resources of anti-stress variation in alfalfa.
Alfalfa callus of Tumu No.2 was used as experimental material,callus was mutated with different concentrations of NaN_3 and different dose of UV,and treated with Na Cl and PEG stress at different concentrations. The growth status of the callus was observed. The semi lethal( LC_(50)) and lethal( LC_(100)) concentration( dose) of each treatment was determined according to the survival rate of callus. The content of proline,soluble sugar,MDA and the activity of CAT in callus were assayed. The results showed that the growth of alfalfa callus was affected by mutation and stress.The survival rate of callus had declined with the increase of NaN_3 concentration and UV irradiation dose.The semi lethal and lethal concentrations of NaN_3 was 4.0 and 5.5×10~(-3) mol·L~(-1) respectively. At the distance of 20 cm,the semi lethal and lethal doses of UV induced mutation was 6 and 8 min. The survival rate of callus declined with the concentration of Na Cl and PEG increasing. The lethal and lethal concentrations of Na Cl was 1.5% and 2% respectively. The lethal and lethal concentrations of PEG was 25% and 30% respectively. Compared with different mutation and stress treatments,the survival rate of callus was 16% treated by NaN_3 and UV combined mutagenesis and PEG and Na Cl double stress treatment. The content of proline,soluble sugar,CAT enzyme increased and MDA content decreased in the callus under NaN_3 and UV combined mutagenesis. Screening of alfalfa callus by mutagenesis and stress could improve the salt and drought resistance. It provides the basis for using somatic cells to expand the resources of anti-stress variation in alfalfa.
引文
[1]安学丽,蔡一林.化学诱变及其在农作物育种上应用[J].核农学报,2003,17(3):239-242.
[2]杨兆民,张璐.辐射诱变技术在农业育种中的应用与探析[J].基因组学与应用生物学,2011,30(1):87-91.
[3]钮力亚,于亮,付晶,等.叠氮化钠在农作物育种中的应用.河北农业科学,2010,14(12):52-53.
[4]彭选明,易卫平,杨震,等.湖南水稻种质资源创制及辐射诱变育种应用成效[J].湖南农业科学,2015,(4):10-13.
[5]吴正景,张菊平,时灿辉,等.叠氮化钠诱变玉扇愈伤组织的研究[J].植物生理学通报,2010,46(12):1247-1250.
[6]卜婷,范小峰,刘丽琼,等.紫外辐射对矮牵牛愈伤组织增殖的影响[J].陇东学院学报,2013,24(5):49-52.
[7]李波,贾秀峰.紫外线诱变苜蓿愈伤组织抗旱性的研究[J].干旱地区农业研究,2005,23(6):103-105.
[8]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.
[9]鹿巍.紫外线及EMS方法诱导小麦叶锈菌毒性突变研究[D].石家庄:河北农业大学,2007.
[10]毛桂莲,郑国琦,戴珍珍.Na Cl和Na2CO3胁迫对枸杞愈伤组织生理效应的影响[J].西北农业学报,2006,15(6):153-157.
[11]Nagy Z,Guoth A,Nemeth E,etal.Metabolic indicators of drought stress Tolerance in wheat:glutaminesynthetase isoenzymes and Rubisco[J].Plant Physiology and Biochemistry,2013,6(7):48-54.
[12]阮先乐,王俊生,陈龙,等.甘薯愈伤组织对盐胁迫的生理反应研究[J].周口师范学院学报,2014,31(5):89-91.
[2]杨兆民,张璐.辐射诱变技术在农业育种中的应用与探析[J].基因组学与应用生物学,2011,30(1):87-91.
[3]钮力亚,于亮,付晶,等.叠氮化钠在农作物育种中的应用.河北农业科学,2010,14(12):52-53.
[4]彭选明,易卫平,杨震,等.湖南水稻种质资源创制及辐射诱变育种应用成效[J].湖南农业科学,2015,(4):10-13.
[5]吴正景,张菊平,时灿辉,等.叠氮化钠诱变玉扇愈伤组织的研究[J].植物生理学通报,2010,46(12):1247-1250.
[6]卜婷,范小峰,刘丽琼,等.紫外辐射对矮牵牛愈伤组织增殖的影响[J].陇东学院学报,2013,24(5):49-52.
[7]李波,贾秀峰.紫外线诱变苜蓿愈伤组织抗旱性的研究[J].干旱地区农业研究,2005,23(6):103-105.
[8]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.
[9]鹿巍.紫外线及EMS方法诱导小麦叶锈菌毒性突变研究[D].石家庄:河北农业大学,2007.
[10]毛桂莲,郑国琦,戴珍珍.Na Cl和Na2CO3胁迫对枸杞愈伤组织生理效应的影响[J].西北农业学报,2006,15(6):153-157.
[11]Nagy Z,Guoth A,Nemeth E,etal.Metabolic indicators of drought stress Tolerance in wheat:glutaminesynthetase isoenzymes and Rubisco[J].Plant Physiology and Biochemistry,2013,6(7):48-54.
[12]阮先乐,王俊生,陈龙,等.甘薯愈伤组织对盐胁迫的生理反应研究[J].周口师范学院学报,2014,31(5):89-91.