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脯氨酸过度积累可缓解盐胁迫并保护转基因高粱[双色高粱(L.)Moench]的光合和抗氧化酶活性。

Proline over-accumulation alleviates salt stress and protects photosynthetic and antioxidant enzyme activities in transgenic sorghum [Sorghum bicolor (L.) Moench].

作者信息

Surender Reddy P, Jogeswar Gadi, Rasineni Girish K, Maheswari M, Reddy Attipalli R, Varshney Rajeev K, Kavi Kishor P B

机构信息

Department of Genetics, Osmania University, Hyderabad 500 007, India.

Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad 500 047, India.

出版信息

Plant Physiol Biochem. 2015 Sep;94:104-13. doi: 10.1016/j.plaphy.2015.05.014. Epub 2015 May 30.

Abstract

Shoot-tip derived callus cultures of Sorghum bicolor were transformed by Agrobacterium tumefaciens as well as by bombardment methods with the mutated pyrroline-5-carboxylate synthetase (P5CSF129A) gene encoding the key enzyme for proline biosynthesis from glutamate. The transgenics were selfed for three generations and T4 plants were examined for 100 mM NaCl stress tolerance in pot conditions. The effect of salt stress on chlorophyll and carotenoid contents, photosynthetic rate, stomatal conductance, internal carbon dioxide concentration, transpiration rates, intrinsic transpiration and water use efficiencies, proline content, MDA levels, and antioxidant enzyme activities were evaluated in 40-day-old transgenic lines and the results were compared with untransformed control plants. The results show that chlorophyll content declines by 65% in untransformed controls compared to 30-38% loss (significant at P < 0.05) in transgenics but not carotenoid levels. Photosynthetic rate (PSII activity) was reduced in untransformed controls almost completely, while it declined by 62-88% in different transgenic lines. Salinity induced ca 100% stomatal closure in untransformed plants, while stomatal conductance was decreased only by 64-81% in transgenics after 4 days. The intercellular CO2 decreased by ca 30% in individual transgenic lines. Malondialdehyde (MDA) content was lower in transgenics compared to untransformed controls. The activities of superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6) and glutathione reductase (GR; EC1.8.1.7) were quantified in leaves exposed to 100 mM NaCl stress and found higher in transgenics. The results suggest that transgenic lines were able to cope better with salt stress than untransformed controls by protecting photosynthetic and antioxidant enzyme activities.

摘要

利用农杆菌介导法以及基因枪轰击法,将编码从谷氨酸合成脯氨酸的关键酶的突变型吡咯啉 - 5 - 羧酸合成酶(P5CSF129A)基因导入双色高粱茎尖来源的愈伤组织培养物中。转基因植株连续自交三代,对T4代植株在盆栽条件下进行100 mM NaCl胁迫耐受性检测。评估了盐胁迫对40日龄转基因株系中叶绿素和类胡萝卜素含量、光合速率、气孔导度、胞间二氧化碳浓度、蒸腾速率、内在蒸腾和水分利用效率、脯氨酸含量、丙二醛(MDA)水平以及抗氧化酶活性的影响,并将结果与未转化的对照植株进行比较。结果表明,与转基因植株30 - 38%的损失(P < 0.05时显著)相比,未转化对照植株的叶绿素含量下降了65%,但类胡萝卜素水平未下降。未转化对照植株的光合速率(PSII活性)几乎完全降低,而不同转基因株系的光合速率下降了62 - 88%。盐胁迫诱导未转化植株气孔几乎完全关闭,而4天后转基因植株的气孔导度仅降低了64 - 81%。各个转基因株系的胞间二氧化碳浓度降低了约30%。与未转化对照相比,转基因植株中丙二醛(MDA)含量较低。对暴露于100 mM NaCl胁迫下的叶片中的超氧化物歧化酶(SOD;EC 1.15.1.1)、过氧化氢酶(CAT;EC 1.11.1.6)和谷胱甘肽还原酶(GR;EC1.8.1.7)活性进行定量分析,发现转基因植株中的这些酶活性较高。结果表明,转基因株系通过保护光合和抗氧化酶活性,比未转化对照更能耐受盐胁迫。

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