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作物学报 ›› 2019, Vol. 45 ›› Issue (8): 1279-1285.doi: 10.3724/SP.J.1006.2019.94001

• 研究简报 • 上一篇    下一篇

不同收获指数甘蓝型油菜β-淀粉酶活性及其基因家族成员的表达分析

靳舒荣1,2,王艳玫1,2,常悦1,2,王月华1,2,李加纳1,2,倪郁1,2,*()   

  1. 1 西南大学农学与生物科技学院, 重庆 400715
    2 西南大学农业科学研究院, 重庆 400715
  • 收稿日期:2019-01-01 接受日期:2019-04-15 出版日期:2019-08-12 网络出版日期:2019-05-08
  • 通讯作者: 倪郁
  • 作者简介:靳舒荣, E-mail:1509512475@qq.com|王艳玫, E-mail: 18113560032@163.com
  • 基金资助:
    本研究由国家自然科学基金项目(31771694);重庆市基础研究与前沿探索项目(cstc2018jcyjAX0263);重庆市基础研究与前沿探索项目(cstc2016jcyjA0170);中央高校基本科研业务费专项资金(XDJK2017B028);国家现代产业技术体系建设专项资助(CARS-12)

Activity and gene family expression of β-amylase in Brassica napus differing in harvest index

JIN Shu-Rong1,2,WANG Yan-Mei1,2,CHANG Yue1,2,WANG Yue-Hua1,2,LI Jia-Na1,2,NI Yu1,2,*()   

  1. 1 College of Agronomy and Biotechnology, Southwest University, Chongqing 400715, China
    2 Academy of Agricultural Sciences, Southwest University, Chongqing 400715, China
  • Received:2019-01-01 Accepted:2019-04-15 Published:2019-08-12 Published online:2019-05-08
  • Contact: Yu NI
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31771694);the Chongqing Basic and Advanced Research Project(cstc2018jcyjAX0263);the Chongqing Basic and Advanced Research Project(cstc2016jcyjA0170);the Fundamental Research Funds for the Central Universities(XDJK2017B028);the China Agriculture Research System(CARS-12)

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摘要：

油菜“源”器官中光合产物向籽粒转移的效率是提高油菜收获指数的关键环节, 而“源”器官中淀粉酶活性影响同化物向籽粒的运输强度。β-淀粉酶(β-amylase, BAM)及其基因家族成员与油菜高收获指数形成之间的关系还不清楚。本研究选择高产高收获指数型、高产低收获指数型、低产低收获指数型3类油菜品种, 在终花期后5、10、15、20、25 d分别取茎杆、叶片、角果皮与种子, 分析β-淀粉酶活性及其基因家族成员的表达水平。结果表明, β-淀粉酶活性在所检测“源”器官中酶活性总体随发育时期增加。高收获指数型油菜叶片、角果皮中的β-淀粉酶活性显著高于低收获指数型油菜。β-淀粉酶基因家族中, BAM1、BAM4与BAM5在油菜茎、叶及角果皮中的表达量总体随发育时期增加。花后25 d时, BAM1与BAM3在高收获指数油菜叶片、角果皮中的表达量显著高于低收获指数油菜。BAM4与BAM5在高收获指数油菜角果皮中的表达量分别于花后15 d与20 d开始显著高于低收获指数油菜。综合分析认为, BAM1和BAM3可能通过促进叶片与角果皮淀粉分解而加强光合产物向籽粒的运输强度; BAM4与BAM5可能主要通过作用于角果皮淀粉分解而调控光合产物向籽粒的运输。BAM4与BAM5也可能参与了油菜种子中淀粉的调控。

关键词: 甘蓝型油菜, 收获指数, β-淀粉酶;, 基因表达

Abstract:

The transferring efficiency of photosynthate from source organs to grains is the key process to increase the harvest index in Brassica napus, and amylase activity in source organs affects the transport intensity of assimilates to grains. The relationship between β-amylase (BAM) and the formation of high harvest index in rapeseed had not been clear. In this study, three different inbred lines, high-yield and high-harvest index rapeseed, high-yield and low-harvest index rapeseed, and low-yield and low-harvest index rapeseed were selected, and stem, leaf, silique pericarp and seed were sampled at 5, 10, 15, 20, and 25 d after the final flowering to analyze the activity of β-amylase and the expression level of its gene family members. The activity of β-amylase increased with the development of source organs. The activity of β-amylase in leaves and silique pericarps of high-harvest index rapeseed was significantly higher than that of low-harvest index rapeseed. In the β-amylase gene family, the expression of BAM1, BAM4, and BAM5 in the stem, leaf and silique pericarp of rapeseed increased with the development of organs. At 25 d after the final flowering, the expression of BAM1 and BAM3 in leaves and silique pericarps of high-harvest index rapeseed was significantly higher than that of low harvest index rapeseed. The expression of BAM4 and BAM5 in the silique pericarps of rapeseed with high harvest index was significantly higher than that of rapeseed with low harvest index at 15 d and 20 d after the final flowering. Taken together, BAM1 and BAM3 may enhance the transport intensity of photosynthate to grains by promoting starch degradation in leaves and silique pericarps in rapeseed with high harvest index, and BAM4 and BAM5 may regulate the transport of photosynthate to grains mainly by acting on starch degradation in silique pericarps. BAM4 and BAM5 may also be involved in the regulation of starch in rapeseed seeds.

Key words: Brassica napus, harvest index, β-amylase;, gene expression

表1

qPCR相关引物"

名称
Name		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)		 扩增片段大小
Size (bp)
BAM1 ATTGGAGCTTTCCAGTGCTATGACA GCCAGTTATTGTAGTGACCAGCATCG 124
BAM3 GATGTGACTCTGTGCCTGTCCTCA CTCCACAAGGTCCCATTCCCA 141
BAM4 ATTCTTCTGGTTGCCCTATCATTAAA GACAGAAACGCTCCACAATCCC 126
BAM5 CAGATCATAGGAGAAGCCAACAAGA GCGTGGCTGTGGTGGTTGTA 101
Actin7 GTGACAATGGAACTGGAATGGTGA GTGCCTAGGACGACCAACAATACTC 92

图1

不同油菜品种的产量与收获指数 HH: 高产高收获指数型油菜; HL: 高产低收获指数型油菜; LL: 低产低收获指数型油菜; HI: 收获指数。图中柱形图上方不同小写字母表示品种间差异显著(P < 0.05)。"

图2

甘蓝型油菜不同发育时期不同组织器官β-淀粉酶活性的动态变化缩写同图1。Abbreviations are the same as those given in Fig. 1."

图3

甘蓝型油菜不同发育时期茎内β-淀粉酶基因家族成员的表达"

图4

甘蓝型油菜不同发育时期叶片β-淀粉酶基因家族成员的表达"

图5

甘蓝型油菜不同发育时期角果皮β-淀粉酶基因家族成员的表达"

图6

甘蓝型油菜不同发育时期种子β-淀粉酶基因家族成员的表达"

[1] Hay R K M . Harvest index: a review of its use in plant breeding and crop physiology. Annu Appl Biol, 1995,126:197-216.
doi: 10.1111/aab.1995.126.issue-1
[2] 袁婺洲, 官春云 . 油菜角果内的淀粉酶活性与有关同化物转运的调控. 湖南师范大学自然科学学报, 1995,18(3):74-79.
Yuan W Z, Guan C Y . Regulation of assimilates transportation by amylase activity in rapeseed pods. J Nat Sci Univ Norm Hunan, 1995,18(3):74-79 (in Chinese with English abstract).
[3] 袁婺洲, 官春云 . 影响油菜收获指数的几个生理因子. 作物学报, 1997,23:580-586.
Yuan W Z, Guan C Y . Harvest index in rapeseed affected by a few physiological factors. Acta Agron Sin, 1997,23:580-586 (in Chinese with English abstract).
[4] Yu T S, Zeeman S C, Thorneycroft D, Fulton D C, Dunstan H, Lue W L, Hegemann B, Tung S Y, Umemoto T, Chapple A, Tsai D L, Wang S M, Smith A M, Chen J, Smith S M . alpha-Amylase is not required for breakdown of transitory starch in Arabidopsis leaves. J Biol Chem, 2005,280:9773-9779.
doi: 10.1074/jbc.M413638200
[5] Weise S E, Kim K S, Stewart R P, Sharkey T D . β-Maltose is the metabolically active anomer of maltose during transitory starch degradation. Plant Physiol, 2005,137:756-761.
doi: 10.1104/pp.104.055996
[6] Scheidig A, Fröhlich A, Schulze S, Lloyd J R, Kossmann J . Down regulation of a chloroplast-targeted β-amylase leads to a starch-excess phenotype in leaves. Plant J, 2002,30:581-591.
doi: 10.1046/j.1365-313X.2002.01317.x
[7] Streb S, Zeeman S C . Starch metabolism in Arabidopsis. Arab Book, 2012,10:e0160.
doi: 10.1199/tab.0160
[8] 申鸽子 . 不同生境下油菜高收获指数的激素平衡与调控. 西南大学硕士学位论文, 重庆, 2016.
Shen G Z . Balance and Regulation of Hormones of High Harvest Index Rapeseed (Brassica napus L.) in Different Environments. MS Thesis of Southwest University, Chongqing, China, 2016 (in Chinese with English abstract).
[9] 李加纳, 卢坤, 荐红举, 梁颖, 陆军花, 彭柳, 申鸽子, 张烨, 张超, 杨博, 张莉 . 油菜收获指数研究进展. 中国油料作物学报, 2018,40:640-648.
Li J N, Lu K, Jian H J, Liang Y, Lu J H, Peng L, Shen G Z, Zhang Y, Zhang C, Yang B, Zhang L . Research advances on harvest index of Brassica napus L. Chin J Oil Crop Sci, 2018,40:640-648 (in Chinese with English abstract).
[10] Allen E J, Morgan D G, Ridgman W I . A physiological analysis of the growth of oilseed rape. J Agric Sci, 1971,77:339-341.
doi: 10.1017/S0021859600024515
[11] Chapman J F, Daniels R W, Scarisbrick D H . Field studies on 14C assimilate fixation and movement in oil-seed rape (B. napus). J Agric Sci, 1984,102:23-31.
[12] Pechan P A, Morgan D G . Defoliation and its effects on pod and seed development in oil seed rape (Brassica napus L.). J Exp Bot, 1985,36:458-468.
[13] Tayo T O, Morgan D G . Factors influencing flower and pod development in oil-seed rape (Brassica napus L.). J Agric Sci, 1979,92:363-373.
[14] Smith S M, Fulton D C, Chia T, Thorneycroft D, Chapple A, Dunstan H, Hylton C, Zeeman S C, Smith A M . Diurnal changes in the transcriptome encoding enzymes of starch metabolism provide evidence for both transcriptional and posttranscriptional regulation of starch metabolism in Arabidopsis leaves. Plant Physiol, 2004,136:2687-2699.
doi: 10.1104/pp.104.044347
[15] Monroe J D, Storm A R . The Arabidopsis β-amylase (BAM) gene family: Diversity of form and function. Plant Sci, 2018,276:163-170.
doi: 10.1016/j.plantsci.2018.08.016
[16] Fulton D C, Stettler M, Mettler T, Vaughan C K, Li J, Francisco P, Gil M, Reinhold H, Eicke S, Messerli G, Dorken G, Halliday K, Smith A M, Smith S M, Zeeman S C . β-amylase 4, a noncatalytic protein required for starch breakdown, acts upstream of three active beta-amylases in Arabidopsis chloroplasts. Plant Cell, 2008,20:1040-1058.
doi: 10.1105/tpc.107.056507
[17] Lao N T, Schoneveld O, Mould R M, Hibberd J M, Gray J C, Kavanagh T A . An Arabidopsis gene encoding a chloroplast- targeted β-amylase. Plant J, 1999,20:519-527.
doi: 10.1046/j.1365-313X.1999.00625.x
[18] Valerio C, Costa A, Marri L, Issakidis-Bourguet E, Pupillo P, Trost P, Sparla F . Thioredoxin-regulated β-amylase (BAM1) triggers diurnal starch degradation in guard cells, and in mesophyll cells under osmotic stress. J Exp Bot, 2011,62:545-555.
doi: 10.1093/jxb/erq288
[19] Horrer D, Flütsch S, Pazmino D, Matthews J S, Thalmann M, Nigro A, Leonhardt N, Lawson T, Santelia D . Blue light induces a distinct starch degradation pathway in guard cells for stomatal opening. Curr Biol, 2016,26:362-370.
doi: 10.1016/j.cub.2015.12.036
[20] Kaplan F, Guy C L . RNA interference of Arabidopsis beta- amylase 8 prevents maltose accumulation upon cold shock and increases sensitivity of PSII photochemical efficiency to freezing stress. Plant J, 2005,44:730-743.
doi: 10.1111/tpj.2005.44.issue-5
[21] Monroe J D, Storm A R, Badley E M, Lehman M D, Platt S M, Saunders L K, Schmitz J M, Torres C E . β-amylase 1 and β-amylase3 are plastidic starch hydrolases in Arabidopsis that seem to be adapted for different thermal, pH, and stress conditions. Plant Physiol, 2014,166:1748-1763.
doi: 10.1104/pp.114.246421
[22] Monroe J D, Breault J S, Pope L E, Torres C E, Gebrejesus T B, Berndsen C E, Storm A R . Arabidopsis β-amylase 2 is a K+-requiring, catalytic tetramer with sigmoidal kinetics . Plant Physiol, 2017,175:1125-1135.
[23] Laby R J, Kim D, Gibson S I . The ram1 mutant of Arabidopsis exhibits severely decreased β-amylase activity. Plant Physiol, 2001,127:1798-1807.
[24] 黄露, 陶诗顺, 张敏, 姜磊, 彭雅利 . 甘蓝型杂交油菜收获指数及其品种间差异性研究. 江苏农业科学, 2011, (1):95-97.
Huang L, Tao S S, Zhang M, Jiang L, Peng Y L . Differences analysis of harvest and varieties of Brassica napus L. Jiangsu Agric Sci, 2011, (1):95-97 (in Chinese with English abstract).
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