Chinese Bulletin of Botany ›› 2017, Vol. 52 ›› Issue (5): 568-578.DOI: 10.11983/CBB16225
• EXPERIMENTAL COMMUNICATIONS • Previous Articles Next Articles
Li Ma1, Wancang Sun1,*(), Jinhai Yuan1, Zigang Liu1, Junyan Wu1, Yan Fang1, Yaozhao Xu2, Yuanyuan Pu1, Jing Bai1, Xiaoyun Dong1, Huili He1
Received:
2016-11-23
Accepted:
2017-03-24
Online:
2017-09-01
Published:
2017-07-10
Contact:
Wancang Sun
Li Ma, Wancang Sun, Jinhai Yuan, Zigang Liu, Junyan Wu, Yan Fang, Yaozhao Xu, Yuanyuan Pu, Jing Bai, Xiaoyun Dong, Huili He. Expression Analysis of β-1,3-Glucanase Gene from Winter Brassica rapa Under Low Temperature Stress[J]. Chinese Bulletin of Botany, 2017, 52(5): 568-578.
Figure 1 Apoplast protein map of Brassica rapa cv. ‘Long- you6’ leavesM: Protein marker; 1: Subtilisin-like protease; 2: Chloroplast heat shock protein 70-1; 3: Myrosinase; 4: S-adenosyl-L- homocystein hydrolase; 5: Basic glucanase; 6: β-1,3-gluca- nase; 7: Pathogenesis- related protein-1
Spot No. | NCBI accession No. | Protein name | Organism | Score | Sequence coverage (%) |
---|---|---|---|---|---|
1 | gi|757534 | Subtilisin-like protease | Arabidopsis thaliana | 146 | 3 |
2 | gi|166919370 | Chloroplast heat shock protein 70-1 | Ipomoea nil | 291 | 6 |
3 | gi|11034734 | Myrosinase | Raphanus sativus | 98 | 6 |
4 | gi|32967699 | S-adenosyl-L-homocystein hydrolase | A. thaliana | 290 | 29 |
5 | gi|118763538 | Basic glucanase | Brassica juncea | 315 | 53 |
6 | gi|62361691 | β-1,3-glucanase | B. rapa subsp. | 348 | 48 |
7 | gi|722274 | Pathogenesis-related protein-1 | B. juncea | 78 | 41 |
Table 1 Mass spectrometry results of apoplast proteins in Brassica rapa cv. ‘Longyou6’ leaves
Spot No. | NCBI accession No. | Protein name | Organism | Score | Sequence coverage (%) |
---|---|---|---|---|---|
1 | gi|757534 | Subtilisin-like protease | Arabidopsis thaliana | 146 | 3 |
2 | gi|166919370 | Chloroplast heat shock protein 70-1 | Ipomoea nil | 291 | 6 |
3 | gi|11034734 | Myrosinase | Raphanus sativus | 98 | 6 |
4 | gi|32967699 | S-adenosyl-L-homocystein hydrolase | A. thaliana | 290 | 29 |
5 | gi|118763538 | Basic glucanase | Brassica juncea | 315 | 53 |
6 | gi|62361691 | β-1,3-glucanase | B. rapa subsp. | 348 | 48 |
7 | gi|722274 | Pathogenesis-related protein-1 | B. juncea | 78 | 41 |
Figure 3 Nucleotide sequence and deduced amino acid sequence of β-1,3-glucanase cDNAAn upstream in-flame start codon ATG is boxed; * indicates the positions of termination codon TAA.
Figure 4 Bioinformatic analysis of β-1,3-glucanase protein in Brassica rapa cv. ‘Longyou6’(A) Predicted protein hydrophobicity by ProtScale; (B) Predicted signal peptide by Signal P4; (C) Transmembrane helical segments predicted by TMpred; (D) Predication of domains; (E) Predicted secondary structure; (F) Predicted 3-D structure
Cruciferous plants | No. of amino acids | Molecular weight (kDa) | Theoretical isoelectric point | Protein instability index | Grand average of hydropathicity | Composition of amino acid | The main amino acid (%) |
---|---|---|---|---|---|---|---|
Longyou6 | 343 | 38.102 | 6.63 | 39.58 | -0.334 | 20 | Asn (9.6), Leu (8.7) |
Tianyou4 | 343 | 38.130 | 7.68 | 39.02 | -0.337 | 20 | Asn (9.6), Leu (8.7) |
Brassica rapa | 342 | 38.180 | 7.69 | 42.99 | -0.370 | 20 | Asn (9.6), Leu (8.8) |
B. rapa subsp. chinensis | 363 | 40.659 | 9.27 | 39.84 | -0.335 | 20 | Asn (8.8), Leu (8.3) |
B. napus | 363 | 40.599 | 9.22 | 39.96 | -0.325 | 20 | Asn (9.1), Leu (8.3) |
B. oleracea | 351 | 38.925 | 6.85 | 40.48 | -0.304 | 20 | Asn (8.8), Ser (8.5) |
B. oleracea var. oleracea | 365 | 41.075 | 9.27 | 37.84 | -0.390 | 20 | Asn (8.8), Leu (7.7) |
B. juncea | 346 | 38.087 | 9.13 | 38.03 | -0.188 | 20 | Gly (9.5), Leu (9.0) |
Raphanus sativus | 345 | 38.243 | 6.20 | 40.33 | -0.326 | 20 | Leu (10.1), Asn (8.4) |
Arabidopsis thaliana | 355 | 39.380 | 5.47 | 47.15 | -0.318 | 20 | Ser (9.3), Leu (8.7) |
Table 2 Predicted physicochemical properties of β-1,3-glucanase gene encoding proteins in cruciferae plants
Cruciferous plants | No. of amino acids | Molecular weight (kDa) | Theoretical isoelectric point | Protein instability index | Grand average of hydropathicity | Composition of amino acid | The main amino acid (%) |
---|---|---|---|---|---|---|---|
Longyou6 | 343 | 38.102 | 6.63 | 39.58 | -0.334 | 20 | Asn (9.6), Leu (8.7) |
Tianyou4 | 343 | 38.130 | 7.68 | 39.02 | -0.337 | 20 | Asn (9.6), Leu (8.7) |
Brassica rapa | 342 | 38.180 | 7.69 | 42.99 | -0.370 | 20 | Asn (9.6), Leu (8.8) |
B. rapa subsp. chinensis | 363 | 40.659 | 9.27 | 39.84 | -0.335 | 20 | Asn (8.8), Leu (8.3) |
B. napus | 363 | 40.599 | 9.22 | 39.96 | -0.325 | 20 | Asn (9.1), Leu (8.3) |
B. oleracea | 351 | 38.925 | 6.85 | 40.48 | -0.304 | 20 | Asn (8.8), Ser (8.5) |
B. oleracea var. oleracea | 365 | 41.075 | 9.27 | 37.84 | -0.390 | 20 | Asn (8.8), Leu (7.7) |
B. juncea | 346 | 38.087 | 9.13 | 38.03 | -0.188 | 20 | Gly (9.5), Leu (9.0) |
Raphanus sativus | 345 | 38.243 | 6.20 | 40.33 | -0.326 | 20 | Leu (10.1), Asn (8.4) |
Arabidopsis thaliana | 355 | 39.380 | 5.47 | 47.15 | -0.318 | 20 | Ser (9.3), Leu (8.7) |
[1] |
陈芳兰, 林玉玲, 陈裕坤, 冯新, 张梓浩, 赖钟雄 (2015). 三明野生蕉β-1,3-葡聚糖酶基因克隆及其低温下SA处理后的表达分析. 西北植物学报 35, 1709-1721.
DOI URL |
[2] |
程红梅, 简桂良, 倪万潮, 杨红华, 王志兴, 孙文姬, 张保龙, 王晓峰, 马存, 贾士荣 (2005). 转几丁质酶和β-1,3-葡聚糖酶基因提高棉花对枯萎病和黄萎病的抗性. 中国农业科学 38, 1160-1166.
DOI URL |
[3] |
邓文星, 张映 (2007). 实时荧光定量PCR技术综述. 生物技术通报(5), 93-95, 103.
DOI URL |
[4] | 高玉龙, 郭旺珍, 王磊, 张天真 (2007). 一个棉花β-1,3-葡聚糖酶基因全长cDNA的克隆与特征分析. 作物学报 33, 1310-1315. |
[5] | 龚束芳, 杨涛, 车代弟 (2010). 抗冻蛋白溶液中冰晶生长行为的研究. 上海交通大学学报(农业科学版) 28, 265-268, 279. |
[6] | 郭尧君 (2005). 蛋白质电泳实验技术(第2版). 北京: 科学出版社. pp. 123-156. |
[7] | 何江峰, 韩冰, 郭慧琴, 张占雄, 赵雅丽 (2007). 燕麦β-1,3-葡聚糖酶II基因3′端cDNA的克隆及分析. 生物技术 17(1), 5-8. |
[8] | 何江峰, 韩冰, 赵宏鑫 (2006). 植物β-1,3-葡聚糖酶的研究进展. 内蒙古农业科技 5, 21-25. |
[9] |
蒋选利, 李振岐, 康振生 (2005). β-1,3-葡聚糖酶与植物的抗病性. 西北农业学报 14, 135-139.
DOI URL |
[10] |
蓝海燕, 王长海, 张丽华, 刘桂珍, 王岚兰, 陈正华, 田颖川 (2000). 导入β-1,3-葡聚糖酶及几丁质酶基因的转基因可育油菜及其抗菌核病的研究. 生物工程学报 16, 142-146.
DOI URL |
[11] |
林植芳, 刘楠 (2012). 活性氧调控植物生长发育的研究进展. 植物学报 47, 74-86.
DOI URL |
[12] |
马文月 (2004). 植物冷害和抗冷性的研究进展. 安徽农业科学 32, 1003-1006.
DOI URL |
[13] |
欧阳波, 李汉霞, 叶志彪 (2002). 植物β-1,3-葡聚糖酶及其基因. 中国生物工程杂志 22, 18-23.
DOI URL |
[14] |
孙万仓, 马卫国, 雷建民, 刘秦, 杨仁义, 武军艳, 王学芳, 叶剑, 曾军, 张亚宏, 康艳丽, 郭秀娟, 魏文惠, 杨杰, 蒲媛媛, 曾潮武, 刘红霞 (2007). 冬油菜在西北旱寒区的适应性和北移的可行性研究. 中国农业科学 40, 2716-2726.
DOI URL |
[15] | 孙万仓, 刘自刚, 周冬梅, 张仁陟 (2016). 北方冬油菜北移与区划. 北京: 科学出版社. pp. 45-47. |
[16] |
杨刚, 史鹏辉, 孙万仓, 刘自刚, 曾秀存, 武军艳, 方彦, 李学才, 陈奇, 刘林波, 杨建胜, 方园, 张娟 (2016). 白菜型冬油菜质外体抗冻蛋白研究. 中国生态农业学报 24, 210-217.
DOI URL |
[17] | 翟国会, 阮小蕾, 吴丽婷, 谭小勇, 李华平 (2011). 指天蕉β-1,3-葡聚糖酶基因全长cDNA的克隆及序列分析. 中国农业科学 44, 3134-3141. |
[18] |
张妙霞 (2010). 野生香蕉(Musa spp., AB group)抗寒相关基因的克隆与表达分析. 博士论文. 福州: 福建农林大学. pp. 134-140.
DOI URL |
[19] | 赵娟, 兰海燕 (2011). 拟南芥β-1,3-葡聚糖酶基因(BG1)在不同组织及非生物胁迫下的表达研究. 新疆农业科学 48, 712-718. |
[20] |
周丽英, 杨丽涛, 郑坚瑜 (2001). 植物抗寒冻基因工程研究进展. 植物学通报 18, 325-331.
DOI URL |
[21] |
Beffa R, Meins Jr FM (1996). Pathogenesis-related function of plant β-1,3-glucanae investigated by antisense transformation—a review.Gene 179, 97-103.
DOI URL PMID |
[22] |
Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochem 72, 248-254.
DOI URL |
[23] |
Griffith M, Ala P, Yang DSC, Hon WC, Moffatt BA (1992). Antifreeze protein produced endogenously in winter rye leaves.Plant Physiol 100, 593-596.
DOI URL |
[24] |
Hon WC, Griffith M, Chong P, Yang DSC (1994). Extraction and isolation of antifreeze proteins from winter rye (Secale cereale L.) leaves.Plant Physiol 104, 971-980.
DOI URL PMID |
[25] |
Masoud SA, Zhu Q, Lamb C, Dixon RA (1996). Constitutive expression of an inducible β-1,3-glucanase in alfalfa reduces disease severity caused by the oomycete patho- gen Phytophthora megasperma f. spmedicaginis, but does not reduce disease severity of chitin-containing fungi.Transgenic Res 5, 313-323.
DOI URL |
[26] |
Mezabasso L, Alberdi M, Raynal M, Ferrerocadinanos ML, Delseny M (1986). Changes in protein synthesis in rapeseed (Brassica napus) seedlings during a low temperature treatment.Plant Physiol 82, 733-738.
DOI URL PMID |
[27] |
Nakamura Y, Sawada H, Kobayashi S, Nakajima I, Yoshikawa M (1999). Expression of soybean β-1,3-endo- glucanase cDNA and effect on disease tolerance in kiwifruit plants.Plant Cell Rep 18, 527-532.
DOI URL |
[28] |
Xiong LM, Zhu JK (2001). Abiotic stress signal transduction in plants: molecular and genetic perspectives.Physiol Plant 112, 152-166.
DOI URL PMID |
[29] |
Zhu Q, Maher EA, Masoud S, Dixon RA, Lamb CJ (1994). Enhanced protection against fungal attack by constitutive co-expression of chitinase and glucanase genes in trans- genic tobacco.Biol Technol 12, 807-812.
DOI URL |
[1] | Yawen Zhang, Shan Liang, Guoyun Xu, Wuxia Guo, Shulin Deng. Genome-wide Identification and Analysis of CONSTANS-like Gene Family in Nicotiana tabacum [J]. Chinese Bulletin of Botany, 2021, 56(1): 33-43. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||