Sun Jie, Dong Yan, Meng Yuemei, Bi Jingran, Liu Hongmei, Ren Junjie, Wang Jinmao, Ren Yachao, Yang Minsheng
Forest Department, College of Forestry, Hebei Agricultural University, Baoding, 071000, China.
Hebei Key Laboratory for Tree Genetic Resources and Forest Protection, Baoding, 071000, China.
Transgenic Res. 2025 Jan 28;34(1):11. doi: 10.1007/s11248-025-00430-3.
To explore the effects of salt-tolerance gene accumulation on salt tolerance in transgenic plant, we used four types of plant expression vector (N27, N28, N29, and N30) carrying mtlD, mtlD + gutD, mtlD + gutD + BADH, mtlD + gutD + BADH + sacB genes respectively, to transform tobacco through Agrobacterium-mediated method. Transgenic lines were identified through polymerase chain reaction (PCR) detection. Transgenic lines and non-transgenic plant (CK) were subjected to 6‰ sodium chloride solution stress; then, fluorescence quantitative PCR (FQ-PCR) and salt tolerance indexes were used to assess characteristics. PCR showed the exogenous genes had been integrated into the tobacco genome. FQ-PCR showed under clean water treatment the target genes were expressed in all transgenic plants at the transcriptional level. The transcript abundances of target genes changed with the number of genes increased, and improved following salt stress. Comparative analyses of salt tolerance indexes showed height growth, biomass (except for N29), chlorophyll content, net photosynthetic rate, Fv/Fm, and PI of all transgenic plants and CK were lower under salt stress than under clean water treatment, to varying degrees. However, the descent ratio was smaller in transgenic plants. A comprehensive evaluation of multiple salt-tolerance indicators performed using the membership function method showed the average salt tolerance of each vector transgenic line was higher than that of CK, and salt tolerance was greater in transgenic polyvalent gene lines than in transgenic monovalent gene lines. The average salt tolerance was N29 > N28 > N30 > N27 > CK. This study provides a theoretical and practical reference for salt tolerance breeding in other plants.
为探究耐盐基因积累对转基因植物耐盐性的影响，我们使用了四种分别携带mtlD、mtlD + gutD、mtlD + gutD + BADH、mtlD + gutD + BADH + sacB基因的植物表达载体（N27、N28、N29和N30），通过农杆菌介导法转化烟草。通过聚合酶链反应（PCR）检测鉴定转基因株系。将转基因株系和非转基因植株（CK）置于6‰氯化钠溶液胁迫下；然后，利用荧光定量PCR（FQ-PCR）和耐盐性指标评估其特性。PCR结果表明外源基因已整合到烟草基因组中。FQ-PCR结果表明，在清水处理下，所有转基因植株中的目标基因在转录水平上均有表达。目标基因的转录丰度随基因数量的增加而变化，并在盐胁迫后提高。耐盐性指标的比较分析表明，在盐胁迫下，所有转基因植株和CK的株高生长、生物量（N29除外）、叶绿素含量、净光合速率、Fv/Fm和PI均低于清水处理，下降程度不同。然而，转基因植株的下降率较小。利用隶属函数法对多个耐盐指标进行综合评价，结果表明各载体转基因株系的平均耐盐性均高于CK，且转基因多价基因株系的耐盐性高于转基因单价基因株系。平均耐盐性为N29＞N28＞N30＞N27＞CK。本研究为其他植物的耐盐性育种提供了理论和实践参考。
