紫雨桦耐盐性及花色苷合成相关基因的表达特性

吕东林; 林琳; 郭译文; 韩锐; 姜静

doi:10.3969/j.issn.1000-2006.201704048

吕东林,林琳,郭译文,韩锐,姜静

南京林业大学学报（自然科学版） ›› 2018, Vol. 42 ›› Issue (02) : 25-32.

PDF(2073194 KB)

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

PDF(2073194 KB)

南京林业大学学报（自然科学版） ›› 2018, Vol. 42 ›› Issue (02) : 25-32. DOI: 10.3969/j.issn.1000-2006.201704048

研究论文

紫雨桦耐盐性及花色苷合成相关基因的表达特性

吕东林,林琳,郭译文,韩锐,姜静^*

作者信息 +

Characterization of gene expression in anthocyanin synthesis and salt tolerance of Betula pendula ‘Purple Rain'

LYU Donglin,LIN Lin,GUO Yiwen, HAN Rui,JIANG Jing^*

Author information +

文章历史 +

摘要

【目的】探讨紫雨桦的耐盐性与花色素苷合成相关基因的相关性,为揭示紫雨桦的耐盐机理提供参考。【方法】以紫雨桦和垂枝桦(对照)为试材,测定盐胁迫下其叶绿素荧光参数及生理指标的变化,同时采用qRT-PCR方法分析紫雨桦和垂枝桦中查耳酮合酶基因(CHS)、二氢黄酮醇还原酶基因(DFR)、花色素苷合成基因及MYB转录因子家族基因的表达特性。【结果】经质量分数0.8%NaCl胁迫12 d后,紫雨桦叶片花青素含量指数显著高于垂枝桦,是垂枝桦的3.4倍,其盐害指数显著低于垂枝桦,胁迫12 d的联苯胺蓝(DAB)和氮蓝四唑(NBT)染色显示,紫雨桦叶片中O^-₂和H₂O₂的累积少于垂枝桦; 在叶绿素荧光参数方面,NaCl胁迫12 d时,紫雨桦的F_v/F_m值仍在正常值范围内,与胁迫0 d比较,其qP和Φ_PS_Ⅱ值的降幅也显著小于垂枝桦(P<0.05),紫雨桦的NPQ较0 d提高了282.29%,升幅显著高于垂枝桦(P<0.01)。 0.8%NaCl胁迫下CHS、DFR花色素苷合成基因及MYB转录因子家族基因的表达特性是:紫雨桦中的BpMYB10、BpCHS2和BpDFR5基因表达规律明显不同于垂枝桦,NaCl胁迫下上述3条基因持续高表达。【结论】富含花青素的紫雨桦能够耐受一定浓度范围的NaCl胁迫,紫雨桦中的BpMYB10、BpCHS2和BpDFR5基因表达规律明显不同于垂枝桦,这些基因可能与花青素的合成密切相关,花色苷可增强紫雨桦清除活性氧自由基的能力,减缓膜质氧化,提高其耐盐性。

Abstract

【Objective】 To understand the mechanism of salt tolerance, we examined a possible relationship between salt tolerance and gene expression for anthocyanin biosynthesis in salt tolerant Betula pendula ‘Purple Rain'.【Methods】 We used Betula pendula ‘Purple Rain' and Betula pendula ‘Roth'(control)to measure chlorophyll fluorescence and other physiological indicators under salt stress. We analyzed gene expression of chalcone synthase(CHS), dihydlroflavonol-4-reductase(DRF), and MYB transcription factor related to anthocyanin biosynthesis using RT-PCR. 【Result】 The results showed that anthocyanin was 3.4 times higher in ‘Purple Rain,' whereas salt damage indexes were significantly lower than those of ‘Roth' after 12 d under 0.8% NaCl stress. The reswlts of DAB and NBT staining showed that accumulation of O^-₂ and H₂O₂ in leaves of ‘Purple Rain' was lower than that of ‘Roth'. The F_v/F_m value for ‘Purple Rain' was within the normal range after 12 d under NaCl stress, and the qP and Φ_PS_Ⅱ values were significantly lower than those of ‘Roth'(P < 0.05). The NPQ was 282.29% and was significantly higher than that of ‘Roth'(P < 0.01). Gene expression of BpCHS2,BpDFR5, and BpMYB10, related to anthocyanin biosynthesis, were continuously up-regulated under NaCl stress. 【Conclusion】 Anthocyanin-rich ‘Purple Rain' can tolerate a certain range of NaCl stress. The BpMYB10, BpCHS2, and BpDFR5 of ‘Purple Rain' exhibited a different expression pattern in comparison with ‘Roth', and this might be closely related to anthocyanin biosynthesis. Anthocyanin can enhance the ability of ‘Purple Rain' to remove reactive oxygen radicals, slow down oxidation of membranes, and improve salt tolerance.

引用本文

EndNote

Ris (Procite)

Bibtex

导出引用

吕东林,林琳,郭译文,韩锐,姜静. 紫雨桦耐盐性及花色苷合成相关基因的表达特性[J]. 南京林业大学学报（自然科学版）. 2018, 42(02): 25-32 https://doi.org/10.3969/j.issn.1000-2006.201704048

LYU Donglin,LIN Lin,GUO Yiwen, HAN Rui,JIANG Jing. Characterization of gene expression in anthocyanin synthesis and salt tolerance of Betula pendula ‘Purple Rain'[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2018, 42(02): 25-32 https://doi.org/10.3969/j.issn.1000-2006.201704048

中图分类号： S722.5

参考文献

[1] 于晓南,张启翔. 彩叶植物多彩形成的研究进展[J]. 园艺学报,2000,27(S1): 533-538. DOI: 10.3321/j.issn:0513-353X.2000.z1.012. YU X N,ZHANG Q X. Review of researches on leaf color changing of color-leafed plants[J]. Acta Horticulturae Sinica,2000,27(S1): 533-538.
[2] 郝峰鸽,李保印,杨立峰,等. 几种彩叶植物生长期色素含量研究[J].华北农学报,2007,22(1): 161-163. DOI: 10.3321/j.issn:1000-7091.2007.01.038. HAO F G,LI B Y,YANG L F,et al. Studies on pigment contents of growing leaves in different color-leafed plants[J]. Acta Agriculturae Boreali-Sinica,2007,22(1): 161-163.
[3] 朱书香,王中华,李彦慧,等. 紫叶矮樱叶片色素理化性质的研究[J]. 河北农业大学学报,2010,33(3): 33-36. DOI: 10.3969/j.issn.1000-1573.2010.03.008. ZHU S X,WANG Z H,LI Y H,et al. Study on the physical-chemical properties of leaf pigment of Prunus ×cistenena ‘Purpleleaf and Chevry' [J]. Journal of Agricultural University of Hebei,2010,33(3): 33-36.
[4] 晁月文,李竞芸,张广辉,等. 彩叶植物呈色机理及其育种研究进展[J]. 江苏林业科技,2008,35(4): 46-48. DOI: 10.3969/j.issn.1001-7380.2008.04.014. CHAO Y W,LI J Y,ZHANG G H,et al. Review on coloration mechanism and breeding of color-leafed plants[J]. Journal of Jiangsu Forestry Science and Technology,2008,35(4): 46-48.
[5] 胡可,韩科厅,戴思兰,等. 环境因子调控植物花青素苷合成及呈色的机理[J]. 植物学报,2010,45(3): 307-317. DOI: 10.3969/j.issn.1674-3466.2010.03.002. HU K,HAN K T,DAI S L,et al. Regulation of plant anthocyanin synthesis and pigmentation by environmental factors[J]. Chinese Bulletin of Botany,2010,45(3): 307-317.
[6] 祝钦泷. 彩叶草(Solenostemon scutellarioides)叶色形成相关的花色素苷生物合成途径的分子调控研[D].重庆: 西南大学,2007. ZHU Q L. Studies on the molecular regulation of the anthocyanin biosynthesis related to leaf color of Solenostemon scutellartoides[D]. Chongqing: Southwest University,2007.
[7] 李保印. 金叶女贞和小叶女贞叶绿体色素的研究[J]. 林业科技,2005,30(3): 63-65. DOI: 10.3969/j.issn.1001-9499.2005.03.024. LI B Y. Study on the chlorophyll content in the leaves of Ligustrum vicaryi and L.quihoui[J]. Forestry Science and Technology,2005,30(3): 63-65.
[8] 王庆菊,胡艳丽,李晓磊,等. 紫叶稠李叶片不同叶序花青苷与化学成分的相关性[J]. 山东农业大学学报(自然科学版),2007,38(4): 557-560. DOI: 10.3969/j.issn.1000-2324.2007.04.014. WANG Q J,HU Y L,LI X L,et al. Correlations of pigments and other chemical components of Prunus Virg in Iana Schubert in different leaf position[J]. Journal of Shandong Agricultural University(Natural Science Edition),2007,38(4): 557-560.
[9] LIN L,MU H Z,JIANG J,et al. Transcriptomic analysis of purple leaf determination in birch gene[J]. Gene,2013,526(2): 251-258. DOI:10.1016/j.gene.2013.05.038.
[10] ZHANG H L,YANG B,LIU J,et al. Analysis of structural genes and key transcription factors related to anthocyanin biosynthesis in potato tubers[J]. Scientia Horticulturae,2017,225: 310-316.DOI:10.1016/j.scienta.2017.07.018.
[11] WANG H X, FAN W J,LI H,et al. Functional characterization of dihydroflavonol-4 reductase in anthocyanin biosynthesis of purple sweet potato underlies the direct evidence of anthocyanins function against abiotic stresses[J]. PLoS ONE,2013,8(11): 1932-6203. DOI: 10.1371/journal.pone.0078484.
[12] KOVINICH N,KAYANJA G,CHANOCA A,et al. Abiotic stresses induce different localizations of anthocyanins in Arabidopsis[J]. Plant Signaling & Behavior,2015,10(7): 1559-2316.DOI: 10.1080/15592324.2015.1027850.
[13] 王敏. SOD1模型化合物调控干旱胁迫下水稻细胞的抗氧化活性[D]. 武汉: 华中师范大学,2011. WANG M. SOD1 model compounds regulate antioxidant activity in cells under drought stress of rice[D]. Wuhan: Central China Normal University,2011.
[14] 刘桂丰,杨传平,蔡智军,等. 转betA基因小黑杨的耐盐性分析及优良转基因株系的选择[J]. 林业科学,2006,42(7): 33-36. DOI:10.3321/j.issn:1001-7488.2006.07.006. LIU G F,YANG C P,CAI Z J,et al. Salt tolerance of betA transgenic Populus simonii×P.nigra and selection for superior transgenic plants[J]. Scientia Silvae Sinicae,2006,42(7): 33-36.
[15] YANG G,CHEN S,JIANG J,et al. Transcriptome analysis reveals the role of BpGH3.5 in rootelongation of Betula platyphylla × B. pendula plant[J]. Cell Tiss Organ Cult,2015,121(3): 605-617. DOI: 10.1007/s11240-015-0731-5.
[16] 谢智华,姜卫兵,韩键,等. 叶片花色素苷对植物光合作用影响的研究进展[J]. 植物生理学报,2011,47(6): 545-550. DOI: 10.13592/j.cnki.ppj.2011.06.010. XIE Z H,JIANG W B,HAN J,et al. Progress of effects of leaf anthocyanins on plant photosynthesis[J]. Plant Physiology Journal,2011,47(6): 545-550.
[17] 刘晓芬,李方,殷学仁,等. 花青苷生物合成转录调控研究进展[J]. 园艺学报,2013,40(11): 2295-2306. DOI: 10.16420/j.issn.0513-353x.2013.11.001. LIU X F,LI F,YIN X R,et al. Recent advances in the transcriptional regulation of anthocyanin biosynthesis[J]. Acta Horticulturae Sinica,2013,40(11): 2295-2306.
[18] 陈静,陈启林,程智慧,等. 花青苷对低温弱光处理的番茄(L.esculentum Mill.)幼苗光合机构的保护作用[J]. 中国农业科学,2007(4): 788-793. DOI: 10.3321/j.issn:0578-1752.2007.04.019. CHEN J,CHEN Q L,CHENG Z H,et al. Protective effect of anthocyanin on photosynthetic apparatus of tomato(L. esculentum Mill.)seedling leaves exposed to low temperature and low irradiance[J]. Scientia Agricultura Sinica,2007(4): 788-793.
[19] TANG X H,ZHAO C L,WEN G S,et al. Physiological mechanism for anthocyanins to strengthen the drought tolerance of plants[J]. Agricultural Science & Technology,2014,15(11): 1935-1941.
[20] 陈晓丽. 过表达IbOr和IbMYB基因甘薯增强抗逆性的生理机制[D]. 陕西: 西北农林科技大学,2015. CHEN X L. Overexpression of ibOr and ibMYB gene confers enhanced tolerance to stress in sweet potato(Ipomoes batats L.)[D]. Shaanxi: Northwest A & F University,2015.
[21] 徐德聪,吕芳德,潘晓杰. 叶绿素荧光分析技术在果树研究中的应用[J]. 经济林研究,2003,21(3): 88-91. DOI: 10.14067/j.cnki.1003-8981.2003.03.029. XU D C,LV F D,PAN X J. Application of chlorophyll fluorescence analysis technique to research of fruit trees[J]. Economic Forest Researches,2003,21(3): 88-91.
[22] 李园园,杨光,韦睿,等. 转TabZIP基因白桦的获得及耐盐性分析[J]. 南京林业大学学报(自然科学版),2013,37(5): 6-12. LI Y Y,YANG G,WEI R,et al. TabZIP transferred Betula platyphylla generation and salt tolerance analysis[J]. Journal of Nanjing Forestry University(Natural Sciences Edition),2013,37(5): 6-12.
[23] 周琦.祝遵凌,施曼.盐胁迫对鹅耳枥生长及生理生化特性的影响[J].南京林业大学学报(自然科学版),2015,39(5):56-60. DOI:10.3969/j.issn.1000-2006.2015.05.011. ZHOU Q, ZHU Z L, SHI M. Effects of salt stress on growth, physiological and biochemical characteristics of Carpinus turczaninowii seedlings[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2015, 39(5): 56-60.
[24] 闫芳,王勤礼,郭有燕,等. NaCl胁迫对祁连山野生黄瑞香叶片光合叶绿素荧光特性的影响[J].西北植物学报,2016,36(6): 1182-1189. DOI: 10.7606/j.issn.1000-4025.2016.06.1182. YAN F,WANG Q L,GUO Y Y,et al. Effect of NaCl stress on the leaf photosynthesis and chlorophyll fluorescence parameters of Daphne giraldii nitsche[J]. Acta Botanica Boreali-Occidentalia Sinica,2016,36(6): 1182-1189.
[25] BOREVITZ J O,XIA Y,BLOUNT J,et al. Activation tagging identifies a conserved MYB regulator of phenylpropanoid biosynthesis[J]. The Plant Cell,2000,12(12): 2383-2394.DOI:10.1105/tpc.12.12.2383.
[26] SARMA A D,SHARMA R. Anthocyanin-DNA copigmentation complex mutual protection against oxidative[J]. Phytochemistry,1999,52(7): 1313-1318. DOI:10.1016/s0031-9422(99)00427-6.
[27] MEHRTENS F,KRANZ H,BEDNAREK P,et al. The Arabidopsis transcription factor MYB 12 is a flavonol-specific regulator of phenylpropanoid biosynthesis[J]. Plant Physiology,2005,138(2): 1083-1096. DOI:10.1104/pp.104.058032.
[28] EUGENIO B,CONCETTA L,YANG Z,et al. Retrotransposons control fruit-specic,cold-dependent accumulation of anthocyanins in blood oranges[J]. Plant Cell,2012,24(3): 1242-1255. DOI:10.1105/tpc.111.095232.
[29] 李亚蒙,王庆菊,沈向. 四种李属彩叶树种枝叶花色苷分布的解剖研究[J]. 山东农业大学学报(自然科学版),2006,37(4): 489-494. DOI: 10.3969/j.issn.1000-2324.2006.04.001. LI Y M,WANG Q J,SHEN X. Pigment distribution in fresh shoot tissues of four colored-leaf Prunus taxa[J]. Journal of Shandong Agricultural University(Natural Science Edition),2006,37(4): 489-494.
[30] 胡晓立,李彦慧,陈东亮,等. 3种李属彩叶植物对NaCl胁迫的生理响应[J]. 西北植物学报,2010,30(2): 370-376. HU X L,LI Y H,CHEN D L,et al. Physiological responses of three colored-leaf species of Prunus under NaCl stress[J]. Acta Botanica Boreali-Occidentalia Sinica,2010,30(2): 370-376.
[31] 李彦慧,李向应,白瑞琴,等. 4种李属彩叶树木对SO₂的抗性[J]. 林业科学,2008,44(2): 28-33. DOI: 10.3321/j.issn:1001-7488.2008.02.005. Li Y H,Li X Y,Bai R Q,et al. Resistance to sulfur dioxide of four colored-leaf species in Prunus[J]. Scientia Silvae Sinicae,2008,44(2): 28-33.
[32] 刘从霞,邓明净,王文凤,等. 5种李属彩叶树木抗寒性研究[J].河北农业大学学报,2007,30(5): 36-39. DOI: 10.3969/j.issn.1000-1573.2007.05.010. LIU C X,DENG M J,WANG W F,et al. The study on cold resistance of five species leaf-colored plants of Prunus[J]. Journal of Agricultural University of Hebei,2007,30(5): 36-39.