| 摘要: |
| 以蛋白核小球藻(Chlorella pyrenoidosa)为研究对象, 首先克隆了热激蛋白70(HSP70)和磷 酸甘油酸激酶(PGK)基因全长序列, 然后研究了0-20μg/mL 水杨酸对高温胁迫下HSP70 和PGK 基 因表达, 以及蛋白核小球藻生长、可溶性糖和蛋白含量的影响。结果获得了蛋白核小球藻HSP70 基 因的cDNA 序列2405 bp, 包括60 bp 的5'-非编码区(UTR)、1959 bp 的开放阅读框(ORF)和386 bp 的3'-UTR 和PGK 基因的cDNA 序列1664 bp, 包括35 bp 的5'-UTR, 1398 bp ORF 和231 bp 3'-UTR. 序列比较和分析表明该蛋白核小球藻HSP70 序列与其它绿藻的同源性高达83%-91%, PGK 为 70%-75%.荧光定量PCR 结果显示随着水杨酸浓度的增加, HSP70 和PGK 的表达量在12h 和24h 均有所增加, 而在10 μg/mL 水杨酸浓度下表达量最大, 12h 时分别为对照组的2.57 倍和1.56 倍, 24h 则为1.71 倍和1.79 倍。1-20μg/mL 水杨酸可促进该藻的比生长速率、可溶性糖和蛋白含量的升高。 本文结果表明一定浓度水杨酸对高温胁迫蛋白核小球藻有缓解作用, 且5 和10 μg/mL 水杨酸的效果 最显著。 |
| 关键词: 蛋白核小球藻 水杨酸 热激蛋白70 磷酸甘油酸激酶 高温胁迫 |
| DOI：10.11693/hyhz20150300082 |
| 分类号: |
| 基金项目:浙江省自然科学基金资助, LY13D060007 号; 宁波市科技攻关项目资助, 2014C10023 号。 |
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| EFFECTS OF SALICYLIC ACID ON GROWTH AND STRESS-RELATED GENES OF CHLORELLA PYRENOIDOSA |
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DING Cong-Cong1,2, XU Nian-Jun1,2, ZHANG Lin1,2, SUN Xue1,2, LI Ya-He1,2
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1.School of Marine Sciences, Ningbo University, Ningbo 315211, China;2.Key Laboratory of Marine Biotechnology of Zhejiang Province, Ningbo 315211, China
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| Abstract: |
| We cloned heat stock protein 70 (HSP70) and phosphoglycerate kinase (PGK) genes, and identified the effects of 0-20.0μg/mL salicylic acid on the gene expression of HSP70 and PGK, algal growth, soluble sugar and protein contents of Chlorella pyrenoidosa under high temperature stress. In this study, we obtained a 2405-bp HSP70 cDNA sequence containing a 60-bp 5'-untranslated region (UTR), a 1959-bp open reading frame (ORF), and a 386-bp 3'-UTR; and a 1664-bp PGK cDNA sequence containing a 35-bp 5'-UTR, 1398-bp ORF, and 231-bp 3'-UTR. Sequence comparison and analysis showed that the homology of HSP70 reached 83%-91% with other green algae, and that of PGK was 70%-75%; both are highly conserved. RT-qPCR showed that with increase in salicylic acid concentration, both expression levels of HSP70 and PGK increased at 12h and 24h, maximum in 10μg/mL salicylic acid. Compared to the control, the expression levels of HSP70 and PGK in 10 μg/mL salicylic acid group at 12h exhibited 2.57-fold and 1.56-fold increase, and 1.71-fold and 1.79-fold at 24h, respectively. Salicylic acid in 1-20μg/mL concentrations could improve the algal specific growth rate, soluble sugar and protein contents. Therefore, salicylic acid in a certain concentrations could alleviate high temperature stress, and the best results were observed in 5 and 10 μg/mL concentrations. |
| Key words: Chlorella pyrenoidosa salicylic acid HSP70 phosphoglycerate kinase (PGK) high temperature stress |
