~(60)Co-γ射线对2种狼尾草属牧草的诱变效应
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摘要
【目的】探究~(60)Co-γ射线对"热研4号"杂交狼尾草及"华南"象草的诱变效应,创制新种质,以推动狼尾草的育种工作。【方法】在前期剂量筛选基础之上,以2种狼尾草品种("热研4号"杂交狼尾草和"华南"象草)种茎为材料,利用~(60)Co-γ射线进行辐射,剂量为30 Gy,剂量率为1 Gy·min~(-1)。将辐射后的种茎与未辐射的种茎栽植于大田中,常规田间管理,1个月后统计成活率。待成活的植株生长成熟后,在2个诱变系群体中分别随机挑选30株进行形态指标的测定,共测定7个指标。随后利用分子标记探究辐射对遗传变异的影响。【结果】2个品种的辐射材料,分别随机选取30株进行形态指标测定,与对照相比,发现"热研4号"诱变系中有28株的株高,21株的茎节数及17株的叶宽有显著或极显著差异。"华南"诱变系中有20株的株高有显著或极显著差异,其他形态指标差异不大。2种牧草的突变体大都表现出株高的降低,因此,生物量也随之减少。还发现一个辐射材料RY4-17,由于分蘖数多于对照导致生物量增加。SSR分子标记研究结果表明,在"热研4号"诱变系中,20对引物的平均多态信息含量为0.245,共扩增出83条带,其中多态性条带52条,诱变系与对照之间的遗传相似系数为0.67—0.89,平均值为0.81,差异位点数为9—27,平均为15.9个,UPGMA聚类结果显示突变体RY4-9和RY4-23与对照遗传距离最远,变异程度最大。在"华南"诱变系中,20对引物的平均多态信息含量为0.394,共扩增出81条带,其中多态性条带65条,诱变系与对照之间的遗传相似系数为0.54—0.86,平均值为0.77,差异位点数为10—38,平均为18.8个,UPGMA聚类结果显示突变体HN-24与对照遗传距离最远,变异程度最大。【结论】适宜剂量的~(60)Go-γ射线辐射可以使2种牧草种茎成活率显著降低,同时可以有效诱导2种牧草在形态学和遗传学水平上发生变异。~(60)Go-γ射线辐射诱变是解决狼尾草因无性繁殖为主而导致遗传资源多样性匮乏的有效途径。
【Objective】 Exploring the mutagenic effects of ~(60)Co-γ-rays on P. purpureum × P. americanurn. cv. Reyan No.4 and P. purpureum Schum. cv. Huanan and creating new germplasm, which aims to promote the process of Pennisetum breeding. 【Method】 Based on the pre-dose screening, using stems of the two species of Pennisetum as materials and irradiated by ~(60)Co-γ-rays and the dose was 30 Gy, the dose rate was 1 Gy·min-1. Then, the irradiated stems were planted in the field and conventional field cultivation management was performed. The survivor rate was counted one month later. After the plants to be survived mature, we randomly selected 30 individual plants from the two species Pennisetum and 7 morphological indicators of them were measured. Then, molecular markers were used to explore the effects of radiation on genetic variation. 【Result】30 individual plants of the two species Pennisetum were selected randomly to measure the morphological indicators, respectively. The heights of 28 mutants, internodes of 21 mutants and leaf breadth of 17 mutants of Reyan No.4 have significant differences at 0.05 or 0.01 levels with the control. The heights of 20 mutants of Huanan have significant differences at 0.05 or 0.01 levels with the control, but there were no significant differences in other morphological indicators with the control. Most of the two species of Pennisetum mutants showed a decrease in plant height, and the biomass also decreased. However, we found that a mutant RY4-17 whose biomass is more than control because it has more tillers. The results of SSR molecular markers indicated that the average PIC of the 20 primers was 0.245 in the Reyan No.4 mutants, and a total of 83 bands were obtained, of which 52 bands were polymorphic. The GS between the mutants and the control was 0.67-0.89, the average was 0.81, the number of different sites was 9-27, with an average of 15.9. The UPGMA cluster showed that the distance between RY4-9, RY4-23 and control was the longest and indicated the mutation degree was greatest. In the Huanan mutants, the average PIC of the 20 primers was 0.394, a total of 81 bands were obtained, of which 65 bands were polymorphic. And the GS between the mutants and the control was 0.54-0.86, the average was 0.77, the number of different sites was 10-38, with an average of 18.8. The UPGMA cluster showed that the distance between HN-24 and control was the longest and indicated the mutation degree was greatest. 【Conclusion】 The appropriate dose of ~(60)Co-γ-rays radiation can significantly reduce the survival rate of the two species Pennisetum, and can effectively induce the mutations in the morphological and genetic. The method is an effective way to solve the lack of genetic resources caused by asexual reproduction of Pennisetum.
【Objective】 Exploring the mutagenic effects of ~(60)Co-γ-rays on P. purpureum × P. americanurn. cv. Reyan No.4 and P. purpureum Schum. cv. Huanan and creating new germplasm, which aims to promote the process of Pennisetum breeding. 【Method】 Based on the pre-dose screening, using stems of the two species of Pennisetum as materials and irradiated by ~(60)Co-γ-rays and the dose was 30 Gy, the dose rate was 1 Gy·min-1. Then, the irradiated stems were planted in the field and conventional field cultivation management was performed. The survivor rate was counted one month later. After the plants to be survived mature, we randomly selected 30 individual plants from the two species Pennisetum and 7 morphological indicators of them were measured. Then, molecular markers were used to explore the effects of radiation on genetic variation. 【Result】30 individual plants of the two species Pennisetum were selected randomly to measure the morphological indicators, respectively. The heights of 28 mutants, internodes of 21 mutants and leaf breadth of 17 mutants of Reyan No.4 have significant differences at 0.05 or 0.01 levels with the control. The heights of 20 mutants of Huanan have significant differences at 0.05 or 0.01 levels with the control, but there were no significant differences in other morphological indicators with the control. Most of the two species of Pennisetum mutants showed a decrease in plant height, and the biomass also decreased. However, we found that a mutant RY4-17 whose biomass is more than control because it has more tillers. The results of SSR molecular markers indicated that the average PIC of the 20 primers was 0.245 in the Reyan No.4 mutants, and a total of 83 bands were obtained, of which 52 bands were polymorphic. The GS between the mutants and the control was 0.67-0.89, the average was 0.81, the number of different sites was 9-27, with an average of 15.9. The UPGMA cluster showed that the distance between RY4-9, RY4-23 and control was the longest and indicated the mutation degree was greatest. In the Huanan mutants, the average PIC of the 20 primers was 0.394, a total of 81 bands were obtained, of which 65 bands were polymorphic. And the GS between the mutants and the control was 0.54-0.86, the average was 0.77, the number of different sites was 10-38, with an average of 18.8. The UPGMA cluster showed that the distance between HN-24 and control was the longest and indicated the mutation degree was greatest. 【Conclusion】 The appropriate dose of ~(60)Co-γ-rays radiation can significantly reduce the survival rate of the two species Pennisetum, and can effectively induce the mutations in the morphological and genetic. The method is an effective way to solve the lack of genetic resources caused by asexual reproduction of Pennisetum.
引文
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[2]郑安俭,杨地龙,叶晓青,顾和平.杂交狼尾草高产栽培技术.种子世界,2003(3):31.ZHENG A J,YANG D L,YE X Q,GU H P.High-yielding cultivation technique of hybrid Pennisetum.Seed World,2003(3):31.(in Chinese)
[3]刘国道,白昌军,王东劲,易克贤,韦家少,何华玄,周家锁.热研4号王草选育.草地学报,2002,10(2):92-96.LIU G D,BAI C J,WANG D J,YI K X,WEI J S,HE H X,ZHOU JS.The selection and utilization of reyan No.4 king grass.Acta Agrestia Sinica,2002,10(2):92-96.(in Chinese)
[4]陆炳章,许慰睽,周春霖,卞同洋.杂交狼尾草在盐渍土区生长特性及其应用.中国草地学报,1990(2):30-40.LU B Z,XU W K,ZHOU C L,BIAN T Y.The growth characteristics and application of Pennisetum hybrid in saline soil area.Chinese Journal of Grassland,1990(2):30-40.(in Chinese)
[5]李茂,字学娟,侯冠彧,周汉林.体外产气法评价5种热带禾本科牧草营养价值.草地学报,2013,21(5):1028-1032.LI M,ZI X J,HOU G Y,ZHOU H L.Nutritional evaluation of five tropical gramineous forages.Acta Agrestia Sinica,2013,21(5):1028-1032.(in Chinese)
[6]荣辉,徐安凯,下条雅敬,邵涛.初次刈割象草青贮发酵品质动态.草地学报,2009,17(4):537-539.RONG H,XU A K,XIA T Y J,SHAO T.Changes in fermentation quality of napiergras harvested the first growth.Acta Agrestia Sinica,2009,17(4):537-539.(in Chinese)
[7]JEWELL W J,CUMMINGS R J,RICHARDS B K.Methane fermentation of energy crops:maximum conversion kinetics and in situ biogas purification.Biomass&Bioenergy.1993,5(3/4):261-278.
[8]STRICKER J A,SMITH D W.Economic Development Through Biomass Systems Integration in Central Florida.Golden,Colorado:National Renewable Energy Laboratory,1995.
[9]WOODARD K R,PRINE G M,BACHREIN S.Solar energy recovery by elephantgrass,energycane,and elephantmillet canopies.Crop Science,1993,33(4):824-830.
[10]SCHANK S C,CHYNOWETH D P,TURICK C E,MENDOZA P E.Napiergrass genotypes and plant parts for biomass energy.Biomass&Bioenergy,1993,4(1):1-7.
[11]温晓娜,简有志,解新明.象草资源的综合开发利用.草业科学,2009,26(9):108-112.WEN X N,JIAN Y Z,XIE X M.The comprehensive exploitation and utilization of Pennisetum purpureum.Pratacultural Science,2009,26(9):108-112.(in Chinese)
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