中国水稻科学 ›› 2020, Vol. 34 ›› Issue (6): 491-498.DOI: 10.16819/j.1001-7216.2020.0316
李孟珠, 王高鹏, 巫月, 任怡, 李刚华, 刘正辉, 丁艳锋, 陈琳*
收稿日期:
2020-04-02
修回日期:
2020-05-27
出版日期:
2020-11-10
发布日期:
2020-11-10
通讯作者:
陈琳
基金资助:
Mengzhu LI, Gaopeng WANG, Yue WU, Yi REN, Ganghua LI, Zhenghui LIU, Yanfeng DING, Lin CHEN*
Received:
2020-04-02
Revised:
2020-05-27
Online:
2020-11-10
Published:
2020-11-10
Contact:
Lin CHEN
摘要: 目的植物蔗糖的跨膜运输主要借助蔗糖转运蛋白(sucrose transporter,SUT)进行。在已鉴定出的5个水稻SUT编码基因中,OsSUT4的作用尚不清楚。方法采用CRISPR/Cas9基因编辑技术构建ossut4缺失变体,出现株高降低、分蘖数增加及产量下降等表型。结果通过进一步分析,发现ossut4突变体水稻叶片蔗糖、淀粉积累,反馈抑制光合速率;因源端光合同化物装载受阻,导致库端籽粒蔗糖供应不足,灌浆延迟。对OsSUT4进行亚细胞和组织化学定位,发现OsSUT4在细胞质膜上,且在萌发种子的胚、胚芽鞘的维管束、小穗颖壳、花药、颖果的糊粉层中都有表达,该现象表明OsSUT4具备转运蔗糖的特征。结论综上所述,OsSUT4在水稻蔗糖源端装载以及籽粒库端卸载等生理过程发挥重要作用。
中图分类号:
李孟珠, 王高鹏, 巫月, 任怡, 李刚华, 刘正辉, 丁艳锋, 陈琳. 水稻蔗糖转运蛋白OsSUT4参与蔗糖转运的功能研究[J]. 中国水稻科学, 2020, 34(6): 491-498.
Mengzhu LI, Gaopeng WANG, Yue WU, Yi REN, Ganghua LI, Zhenghui LIU, Yanfeng DING, Lin CHEN. Function Analysis of Sucrose Transporter OsSUT4 in Sucrose Transport in Rice[J]. Chinese Journal OF Rice Science, 2020, 34(6): 491-498.
W= | A | ; 1) | |
---|---|---|---|
(1+Be-kt)1/N | |||
对方程(1)求导,得到灌浆速率R: | |||
R= | AkBe-kt | ; 2) | |
N(1+Be-kt)(N+1)/N |
W= | A | ; 1) | |
---|---|---|---|
(1+Be-kt)1/N | |||
对方程(1)求导,得到灌浆速率R: | |||
R= | AkBe-kt | ; 2) | |
N(1+Be-kt)(N+1)/N |
基因名称 Gene name | 正向引物 Forward primer (5'→3') | 反向引物 Reverse primer(5'→3') |
---|---|---|
Actin | CAATCGTGAGAAGATGACCC | GTCCATCAGGAAGCTCGTAGC |
OsSUT1 | GCTTTCAACCAGGGTGTCAG | ACTTTCCGGCACATTGGTTC |
OsSUT3 | ATCTGTGATCGTCATCGGGTTC | GCACGGTGTTGTTGGAGAAGT |
OsSUT4 | CTCGTGCCCTTTTAGCTGAC | AACGTTTCCAACAGCCATCC |
OsSUT5 | TTCGCCTTCATCTGTGGAGT | GCCTTTGATGGAGTTTCGCA |
表1 荧光定量PCR引物序列
Table 1 Primer sequences of RT-PCR.
基因名称 Gene name | 正向引物 Forward primer (5'→3') | 反向引物 Reverse primer(5'→3') |
---|---|---|
Actin | CAATCGTGAGAAGATGACCC | GTCCATCAGGAAGCTCGTAGC |
OsSUT1 | GCTTTCAACCAGGGTGTCAG | ACTTTCCGGCACATTGGTTC |
OsSUT3 | ATCTGTGATCGTCATCGGGTTC | GCACGGTGTTGTTGGAGAAGT |
OsSUT4 | CTCGTGCCCTTTTAGCTGAC | AACGTTTCCAACAGCCATCC |
OsSUT5 | TTCGCCTTCATCTGTGGAGT | GCCTTTGATGGAGTTTCGCA |
株系名称 Name of line | 株高 Plant height | 分蘖数 Tiller number | 每穗粒数 Grain number per panicle | 结实率 Seed setting rate/% | 一次枝梗数 Number of primary rachis branches | 二次枝梗数 Number of secondary rachis branches | 千粒重 1000-grain weight |
---|---|---|---|---|---|---|---|
WT | 87.5 a | 12.2 b | 87.6 a | 84.0a | 11.0a | 10.0a | 25.6a |
sut4-15-1 | 82.4bc | 12.4 a | 66.9 b | 73.3 b | 8.8b | 7.4c | 24.7 b |
sut4-15-2 | 81.8 c | 12.2 a | 68.4 b | 65.8 c | 8.9 b | 7.8 c | 24.8b |
sut4-46 | 83.3 b | 12.0 a | 68.5 b | 72.8b | 8.8b | 8.9b | 23.5 c |
表2 OsSUT4突变对株高和产量构成的影响
Table 2 Plant height and yield components ofossut4 mutant and wild type.
株系名称 Name of line | 株高 Plant height | 分蘖数 Tiller number | 每穗粒数 Grain number per panicle | 结实率 Seed setting rate/% | 一次枝梗数 Number of primary rachis branches | 二次枝梗数 Number of secondary rachis branches | 千粒重 1000-grain weight |
---|---|---|---|---|---|---|---|
WT | 87.5 a | 12.2 b | 87.6 a | 84.0a | 11.0a | 10.0a | 25.6a |
sut4-15-1 | 82.4bc | 12.4 a | 66.9 b | 73.3 b | 8.8b | 7.4c | 24.7 b |
sut4-15-2 | 81.8 c | 12.2 a | 68.4 b | 65.8 c | 8.9 b | 7.8 c | 24.8b |
sut4-46 | 83.3 b | 12.0 a | 68.5 b | 72.8b | 8.8b | 8.9b | 23.5 c |
图4 OsSUT4突变对水稻蔗糖含量、淀粉含量、净光合速率和SPAD值的影响 A-白天结束和夜晚结束时蔗糖含量;B-白天结束和夜晚结束时淀粉含量;C-净光合速率;D-SPAD值。平均值±标准误(n=3)。不同字母代表株系之间差异达0.05显著水平(P<0.05, 邓肯检验)。
Fig.4. Effects ofOsSUT4mutation on sucrose, starch contents, photosynthetic rate and SPAD value in rice leaves. A, Sucrose content at the end of day and night; B, Starch content at the end of day and night; C, Net photosynthetic rate; D, SPAD value. Values are Mean±SE (n=3). Different letters above bars indicate significant difference(P<0.05) among the lines (Duncan’s test).
图5 OsSUT4突变对水稻花粉活力的影响 A-花粉染色;B-花粉可染率。平均值±标准误(n=5)。不同字母代表株系之间差异达0.05显著水平(P<0.05, 邓肯检验)。
Fig. 5. Effects of OsSUT4 mutation on pollen vigor in rice. A, Pollen staining; B, Pollen dyeability rate. Values are Mean±SE(n=5). Different letters above bars indicate significant difference(P<0.05) among the lines(Duncan’s test).
图7 水稻蔗糖转运蛋白OsSUT4的亚细胞定位(A-D)和组织定位(E-K) A-GFP绿色荧光;B-明场;C-质膜标记OSMCA1-RFP;D-融合荧光;E-萌发的种子;F-叶片;G-叶鞘;H-根;I、J-小穗;K-颖果。
Fig.7. Sub-cellular localization(A-D) of OsSUT4 and tissue expression(E-K) of pOsSUT4:GUS transgenic rice. A, cDNA-GFP; B, Bright field; C, Membrane marker OSMCA1-RFP; D, Merged; E, Germinated seed; F, Leaf; G, Leaf sheath; H, Root; I&J, Spikelet; K, Caryopsis.
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