栽培大豆和滩涂野大豆杂交后代JB185的耐盐生理与分子机理
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摘要
以栽培大豆Jackson品种(耐盐性较弱)和滩涂野生大豆BB52种群(耐盐性较强)及其经逐代耐盐性筛选的杂交后代JB185(F_5)幼苗为材料,用0、50、100、150 mmol·L~(-1)NaCl溶液处理6d,从含水量、根冠比、叶绿素及其荧光参数、盐离子吸收及其毒害等生理方面和通过EST(表达序列标签)方法从分子水平对其耐盐能力或特性进行了比较研究。主要结果如下:
胁迫6d后,供试3材料的相对生长速率、植株含水量、干物质积累量均随着NaCl浓度的增加而下降,根冠比、丙二醛含量、相对电解质渗漏率随着NaCl浓度的增加而上升,其中JB185各项生理指标的变化介于两亲本之间。
在50、100 mmol·L~(-1)NaCl处理6d时,Jackson和JB185幼苗叶片的Chl和Car含量与对照相比上升,在150 mmol·L~(-1)NaCl处理下与对照相比显著下降,而在各NaCl浓度处理下BB52叶片Chl和Car含量与对照相比都表现明显升高趋势。随着NaCl浓度增加,BB52和JB185幼苗叶片Fv/Fm值与对照相比无明显变化,Jackson在50、100 mmol·L~(-1)NaCl处理下Fv/Fm值与对照相比上升,在150mmol·L~(-1)NaCl胁迫下显著下降。
胁迫6d后,除在50 mmol·L~(-1)NaCl胁迫下BB52幼苗根部K~+吸收增强外,3材料无论是不同器官还是植株整体水平上K~+含量随着NaCl浓度的增加均呈下降趋势,BB52体内K~+含量的降幅较Jackson小。Na~+含量随着NaCl浓度的增加均呈上升趋势,在根、茎及植株整体水平上BB52较Jackson Na~+含量均少,但在叶中相反;Na~+/K~+值与Na~+的变化趋势一致。JB185幼苗体内Na~+、K~+含量及Na~+/K~+值均介于两亲本之间。
根据EST数据库拼接并从大豆cDNA中获得了1条大豆Cl~-通道基因的EST序列,通过Southern blot、半定量Northern blot、RT-PCR和分光光度法等检测结果表明,该EST所在Cl~-通道基因在供试3材料中的拷贝数无差异,但在盐胁迫下,栽培大豆Jackson幼苗根和叶中该基因的转录水平表现减弱趋势,而野生大豆BB52和杂交后代JB185表现增强趋势或相对较高水平,在JB185中更为明显。盐胁迫下Jackson幼苗体内积累的Cl~-相对较多,且向叶片运输比例较大,而BB52和JB185体内积累的Cl~-相对较少,且主要积累在根部,向叶片运输比例较小。
上述研究结果表明:相对于其母本栽培大豆Jackson品种而言,通过与滩涂野大豆BB52种群的杂交和逐代耐盐性筛选,其后代JB185(F_5)的耐盐性得以提高,并且可能与Cl-通道基因的转录增强有关。
The Glycine max Jackson cultivar(the salt sensitive),the salt-born Glycine soja BB52 population(the salt tolerant),and their hybrid JB185(F_5) selected for salt tolerance generation by generation were used as the experimental materials.The above seedlings were stressed for 6 d with 50,100,150 mmol·L~(-1) NaCl solution,respectively. The physiological changes in water content,value of R/T,cholophyll and its fluorescence parameter,salt ion absorption and its toxicity,and molecular mechanism mainly through by EST(expressed sequence tags) were investigated.The result are as follows:
After stressed for 6 d,the plant relative growth rate,water content,and dry weight of the 3 experimental materials were declined with the increase of NaC1 concentration. The value of R/T,MDA content,and relative electrolytic leakage were increased with the increase of NaCI concentration.The changes in above physiological indexes of JB185 were among its parents.
The chlorophyll and carotenoid contents in leaves of Jackson and JB185 were increased under 50,100 mmol·L~(-1) NaCl stress,but they were decreased under 150 mmol·L~(-1) NaCl stress when compared to the control.Chlorophyll and carotenoid contents in leaves of BB52 were all increased under these NaC1 stresses.The Fv/Fm values in leaves of BB52 and JB185 were not obviously changed with the increase of NaCl concentration when compared to the control,and those in Jackson were increased under 50,100 mmol·L~(-1) NaCl stresses,but it was obviously decreased under 150 mmol·L~(-1) NaCl stress.
The K~+ contents in different organs or whole seedlings of 3 experimental materials were decreased with the increase of NaCl concentration,except for the enhanced K~+ absorption in roots of BB52 seedlings under 50 mmol·L~(-1)NaCl stress,but the decrease of K~+ content in BB52 was smaller than those in Jackson.The Na~+ contents in the 3 experimental materials at different organs or whole plants were increased with the increase of NaCI concentration.The Na~+ contents in root,stem,and whole seedling of BB52 were all fewer than Jackson,but it showed the reverse in leaf. The change in value of Na~+/K~+ was consistent with the change in Na~+.The Na~+ and K+ contents,and value of Na~+/K~+ in JB 185 seedlings were between its parents.
One EST(expressed sequence tag) of chloride channel genes(CLCs) in soybean was assembled according to the EST database,and was confirmed in soybean cDNA by RT-PCR technique.No difference in gene copy number of the EST among the Glycine max(Jackson),Glycine soja(BB52) and their hybrid(JB185) was showed by Southern blot, its weakening transcriptional level by semi-quantitative Northern blot and RT-PCR exhibited in roots and leaves of Jackson seedlings under salt stress,while an enhanced or relatively higher trend was found in salt-stressed BB52 and JB185 seedlings,especially in JB185.Measured by photometric method,the higher CI content in plants and its larger proportion in leaves of Jackson seedlings,and lower Cl~content in plants and its larger proportion in roots in BB52 and JB185 seedlings were observed.
The results of this research indicate that,the relative improved salt tolerance of the hybrid JB185 compared to its female parent Jackson can be achieved by hybridization with the salt-born BB52 population and selection for salt tolerance generation by generation,and this improved salt tolerance may be resulted from the enhanced transcriptional level of CLCs.
胁迫6d后,供试3材料的相对生长速率、植株含水量、干物质积累量均随着NaCl浓度的增加而下降,根冠比、丙二醛含量、相对电解质渗漏率随着NaCl浓度的增加而上升,其中JB185各项生理指标的变化介于两亲本之间。
在50、100 mmol·L~(-1)NaCl处理6d时,Jackson和JB185幼苗叶片的Chl和Car含量与对照相比上升,在150 mmol·L~(-1)NaCl处理下与对照相比显著下降,而在各NaCl浓度处理下BB52叶片Chl和Car含量与对照相比都表现明显升高趋势。随着NaCl浓度增加,BB52和JB185幼苗叶片Fv/Fm值与对照相比无明显变化,Jackson在50、100 mmol·L~(-1)NaCl处理下Fv/Fm值与对照相比上升,在150mmol·L~(-1)NaCl胁迫下显著下降。
胁迫6d后,除在50 mmol·L~(-1)NaCl胁迫下BB52幼苗根部K~+吸收增强外,3材料无论是不同器官还是植株整体水平上K~+含量随着NaCl浓度的增加均呈下降趋势,BB52体内K~+含量的降幅较Jackson小。Na~+含量随着NaCl浓度的增加均呈上升趋势,在根、茎及植株整体水平上BB52较Jackson Na~+含量均少,但在叶中相反;Na~+/K~+值与Na~+的变化趋势一致。JB185幼苗体内Na~+、K~+含量及Na~+/K~+值均介于两亲本之间。
根据EST数据库拼接并从大豆cDNA中获得了1条大豆Cl~-通道基因的EST序列,通过Southern blot、半定量Northern blot、RT-PCR和分光光度法等检测结果表明,该EST所在Cl~-通道基因在供试3材料中的拷贝数无差异,但在盐胁迫下,栽培大豆Jackson幼苗根和叶中该基因的转录水平表现减弱趋势,而野生大豆BB52和杂交后代JB185表现增强趋势或相对较高水平,在JB185中更为明显。盐胁迫下Jackson幼苗体内积累的Cl~-相对较多,且向叶片运输比例较大,而BB52和JB185体内积累的Cl~-相对较少,且主要积累在根部,向叶片运输比例较小。
上述研究结果表明:相对于其母本栽培大豆Jackson品种而言,通过与滩涂野大豆BB52种群的杂交和逐代耐盐性筛选,其后代JB185(F_5)的耐盐性得以提高,并且可能与Cl-通道基因的转录增强有关。
The Glycine max Jackson cultivar(the salt sensitive),the salt-born Glycine soja BB52 population(the salt tolerant),and their hybrid JB185(F_5) selected for salt tolerance generation by generation were used as the experimental materials.The above seedlings were stressed for 6 d with 50,100,150 mmol·L~(-1) NaCl solution,respectively. The physiological changes in water content,value of R/T,cholophyll and its fluorescence parameter,salt ion absorption and its toxicity,and molecular mechanism mainly through by EST(expressed sequence tags) were investigated.The result are as follows:
After stressed for 6 d,the plant relative growth rate,water content,and dry weight of the 3 experimental materials were declined with the increase of NaC1 concentration. The value of R/T,MDA content,and relative electrolytic leakage were increased with the increase of NaCI concentration.The changes in above physiological indexes of JB185 were among its parents.
The chlorophyll and carotenoid contents in leaves of Jackson and JB185 were increased under 50,100 mmol·L~(-1) NaCl stress,but they were decreased under 150 mmol·L~(-1) NaCl stress when compared to the control.Chlorophyll and carotenoid contents in leaves of BB52 were all increased under these NaC1 stresses.The Fv/Fm values in leaves of BB52 and JB185 were not obviously changed with the increase of NaCl concentration when compared to the control,and those in Jackson were increased under 50,100 mmol·L~(-1) NaCl stresses,but it was obviously decreased under 150 mmol·L~(-1) NaCl stress.
The K~+ contents in different organs or whole seedlings of 3 experimental materials were decreased with the increase of NaCl concentration,except for the enhanced K~+ absorption in roots of BB52 seedlings under 50 mmol·L~(-1)NaCl stress,but the decrease of K~+ content in BB52 was smaller than those in Jackson.The Na~+ contents in the 3 experimental materials at different organs or whole plants were increased with the increase of NaCI concentration.The Na~+ contents in root,stem,and whole seedling of BB52 were all fewer than Jackson,but it showed the reverse in leaf. The change in value of Na~+/K~+ was consistent with the change in Na~+.The Na~+ and K+ contents,and value of Na~+/K~+ in JB 185 seedlings were between its parents.
One EST(expressed sequence tag) of chloride channel genes(CLCs) in soybean was assembled according to the EST database,and was confirmed in soybean cDNA by RT-PCR technique.No difference in gene copy number of the EST among the Glycine max(Jackson),Glycine soja(BB52) and their hybrid(JB185) was showed by Southern blot, its weakening transcriptional level by semi-quantitative Northern blot and RT-PCR exhibited in roots and leaves of Jackson seedlings under salt stress,while an enhanced or relatively higher trend was found in salt-stressed BB52 and JB185 seedlings,especially in JB185.Measured by photometric method,the higher CI content in plants and its larger proportion in leaves of Jackson seedlings,and lower Cl~content in plants and its larger proportion in roots in BB52 and JB185 seedlings were observed.
The results of this research indicate that,the relative improved salt tolerance of the hybrid JB185 compared to its female parent Jackson can be achieved by hybridization with the salt-born BB52 population and selection for salt tolerance generation by generation,and this improved salt tolerance may be resulted from the enhanced transcriptional level of CLCs.
引文
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