Li Kailong, Qiu Hong, Zhou Min, Lin Yang, Guo Zhenfei, Lu Shaoyun
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Grassland Science Engineering Research Center, College of Life Sciences, South China Agricultural University, Guangzhou, China.
College of Grassland Science, Nanjing Agricultural University, Nanjing, China.
Front Plant Sci. 2018 Sep 6;9:1319. doi: 10.3389/fpls.2018.01319. eCollection 2018.
(Aublet) Sw. is a tropical forage legume with soil acidity tolerance and excellent adaptation to infertile soils, but sensitive to chilling. To understand the molecular responses of to chilling, differentially expressed genes between a chilling tolerant mutant 7-1 and the wild type were identified using suppression subtractive hybridization, and eight of them were confirmed and the regulation pattern were analyzed using quantitative reverse transcription PCR (qRT-PCR). Chloroplast protein 12 (CP12) functions to regulate the Calvin cycle by forming a ternary complex with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK). transcript was induced by chilling in both plants, and higher levels were observed in 7-1 than in the wild type, implying a potential role of in chilling tolerance. To confirm this, transgenic plants over-expressing or down-regulating were generated, respectively. Higher / and survival rate and lower ion leakage were observed in transgenic plants overexpressing as compared with the wild type after chilling treatment, while lower / and survival rate and higher ion leakage were found in antisense plants. overexpression plants showed promoted growth with increased plant height and fresh weight, while the antisense plants exhibited reduced growth with decreased plant height and fresh weight as compared with the wild type. The results indicated that regulation of expression alters plant growth and chilling tolerance in . In addition, higher levels of net photosynthetic rate (), GAPDH and PRK activities were observed in overexpression transgenic plants, while lower levels in antisense plants than in the wild type under both control and chilling conditions, indicating that altered activities of GAPDH and PRK were associated with the changed in transgenic . Our results suggest that SgCP12 regulates GAPDH and PRK activities, , and chilling tolerance in .
(奥布莱特)SW是一种热带饲用豆科植物,耐土壤酸性,对贫瘠土壤适应性良好,但对低温敏感。为了解其对低温的分子响应,利用抑制性消减杂交技术鉴定了耐低温突变体7-1与野生型之间的差异表达基因,其中8个基因得到确认,并通过定量逆转录PCR(qRT-PCR)分析了其调控模式。叶绿体蛋白12(CP12)通过与甘油醛-3-磷酸脱氢酶(GAPDH)和磷酸核酮糖激酶(PRK)形成三元复合物来调节卡尔文循环。两种植株中CP12转录本均受低温诱导,且在7-1中的水平高于野生型,这意味着CP12在耐低温方面可能发挥作用。为证实这一点,分别构建了过表达或下调CP12的转基因SW植株。低温处理后,与野生型相比,过表达CP12的转基因植株具有更高的CP12/和存活率以及更低的离子渗漏率,而反义植株则具有更低的CP12/和存活率以及更高的离子渗漏率。过表达CP12的植株表现出促进生长,株高和鲜重增加,而反义植株与野生型相比则表现出生长受抑制,株高和鲜重降低。结果表明,CP12表达的调控改变了SW的植株生长和耐低温性。此外,在对照和低温条件下,过表达转基因植株的净光合速率()、GAPDH和PRK活性均高于野生型,而反义植株则低于野生型,这表明GAPDH和PRK活性的改变与转基因SW中CP12的变化有关。我们的结果表明,SgCP12调节GAPDH和PRK活性、净光合速率以及SW的耐低温性。