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作物学报 ›› 2022, Vol. 48 ›› Issue (9): 2377-2389.doi: 10.3724/SP.J.1006.2022.12057

• 耕作栽培·生理生化 • 上一篇    下一篇

离体饲养下HgCl2影响水稻叶片光合特性及其生理机制研究

王权1(), 王乐乐1, 朱铁忠1, 任浩杰1, 王辉1, 陈婷婷1, 金萍1, 武立权1,2,*(), 杨茹1, 尤翠翠1, 柯健1, 何海兵1,*()   

  1. 1.安徽农业大学农学院, 安徽合肥 230036
    2.江苏省现代作物生产协同创新中心, 江苏南京 210095
  • 收稿日期:2021-08-17 接受日期:2022-01-05 出版日期:2022-09-12 网络出版日期:2022-07-15
  • 通讯作者: 武立权,何海兵
  • 作者简介:E-mail: 1715994244@qq.com
  • 基金资助:
    国家自然科学基金项目(32071946);国家自然科学基金项目(31801286);安徽省自然科学基金项目(1908085MC67);国家级大学生创新创业项目(S202010364129);安徽农业大学校级大创项目(202110364687);安徽农业大学校级大创项目(202110364345)

Effects of HgCl2 on photosynthetic characteristics and its physiological mechanism of rice leaves in vitro feeding

WANG Quan1(), WANG Le-Le1, ZHU Tie-Zhong1, REN Hao-Jie1, WANG Hui1, CHEN Ting-Ting1, JIN Ping1, WU LI-Quan1,2,*(), YANG Ru1, YOU Cui-Cui1, KE Jian1, HE Hai-Bing1,*()   

  1. 1. College of Agronomy, Anhui Agricultural University, Hefei 230036, Anhui, China
    2. Jiangsu Collaborative Center for Modern Crop Production, Nanjing 210095, Jiangsu, China
  • Received:2021-08-17 Accepted:2022-01-05 Published:2022-09-12 Published online:2022-07-15
  • Contact: WU LI-Quan,HE Hai-Bing
  • Supported by:
    National Natural Science Foundation of China(32071946);National Natural Science Foundation of China(31801286);Natural Science Foundation of Anhui Province(1908085MC67);National Student Innovation and Entrepreneurship Project(S202010364129);University-level Large Innovation Project of Anhui Agricultural University(202110364687);University-level Large Innovation Project of Anhui Agricultural University(202110364345)

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摘要：

HgCl2作为水通道蛋白专用抑制剂, 可有效量化水通道蛋白和叶片结构对光合作用的贡献度, 对认识提高作物光合作用的潜在途径具有重要意义。然而, 有关抑制叶片水通道蛋白的HgCl2浓度和时长尚不清楚。本研究以水稻品种Y两优900和徽两优898为试材, 采用HgCl2溶液离体饲养水稻叶片(叶龄余数为2), 设置不同处理浓度: 0、100、200、300和500 µmol L-1及不同处理时间: 0、0.5、1、1.5、2、2.5、3、3.5、4和4.5 h。研究表明, 不同浓度和处理时长对叶片相对含水量无显著影响(P>0.05)。随着浓度增加, 叶片SPAD值、净光合速率和气孔导度呈明显降低趋势。与对照相比, 浓度为100 µmol L-1时长2 h时的净光合速率降低至最低值(62.33%), 随着处理时间延长, 光合速率趋于稳定。当浓度>100 µmol L-1时, 随着处理时长增加, 净光合速率持续降低, 且超氧化物歧化酶(SOD)和过氧化物酶(POD)活性及丙二醛(MDA)含量均显著增加(P<0.05), 表明浓度>100 µmol L-1, 处理时间较长时, HgCl2溶液能对叶片活体造成伤害。与活体测量相比, 叶片净光合速率离体稳定数值降幅约15%~20%, 因而离体叶片测量光合速率乘以1.25~1.33的矫正系数, 可能较准确反映水稻叶片活体原位测定的光合指标。此外, 100 µmol L-1 HgCl2显著降低水稻叶片水通道蛋白基因表达。这些结果表明, HgCl2高效安全抑制水稻水通道蛋白的最佳组合为100 µmol L-1下饲养2 h。

关键词: 水稻, HgCl2, 离体饲养, 光合作用, 抗氧化酶活性

Abstract:

As a special inhibitor of aquaporin, HgCl2 can effectively quantify the contribution of aquaporin and leaf structure to photosynthesis, which is of great significance for understanding the potential ways to improve crop photosynthesis. However, the concentration and duration of HgCl2 inhibiting aquaporin in leaves are still unclear. In this study, rice varieties Y liangyou 900 and Huiliangyou 898 were used as materials to feed rice leaves in vitro with HgCl2 solution (the remainder of leaf age was 2), and different treatment concentrations (0, 100, 200, 300, and 500 µmol L-1), and different treatment times (0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, and 4.5 h) were set. The results showed that different concentrations and treatment times had no significant effect on the relative water content of leaves (P > 0.05). With the increase of concentration, the SPAD value, net photosynthetic rate, and stomatal conductance of leaves decreased obviously. Compared with the control, the net photosynthetic rate decreased to the lowest value (62.33%) when the concentration was 100 µmol L-1 for 2 h, and the net photosynthetic rate tended to be stable with the extension of treatment time. When the concentration was less than 100 µmol L-1, the net photosynthetic rate decreased continuously with the increase of treatment time, and the activities of superoxide dismutase (SOD), peroxidase (POD), and malondialdehyde content increased significantly (P < 0.05), indicating that HgCl2 solution could damage the living leaves when the concentration was less than 100 µmol L-1 and the treatment time was longer. Compared with in vivo measurement, the steady value of net photosynthetic rate of leaves in vitro decreased by about 15%-20%, so the correction coefficient of measured net photosynthetic rate of leaves in vitro multiplied by 1.25-1.33 might accurately reflect the photosynthetic index of rice leaves in vivo measurement. In addition, 100 µmol L-1 HgCl2 significantly reduced the expression of aquaporin gene in rice leaves. Therefore, the optimal combination of HgCl2 to effectively and safely inhibit aquaporin in rice was 100 µmol L-1 for 2 h.

Key words: rice, HgCl2, in vitro feeding, photosynthesis, antioxidant enzyme activity

图1

HgCl2离体饲养处理及水稻叶片光合气体交换参数获取"

表1

水稻水通道蛋白基因(OsPIPs)名称及引物序列"

基因序列号
Gene ID		 引物名称
Primer name		 序列
Sequence (5'-3')
Os02g0666200 OsPIP1-1F TGAGATTGTTGGCACCTTCA
OsPIP1-1R AGTGGGGCAAGGATAGGAAC
Os04g0559700 OsPIP1-2F ACCAGGGCCCTCTTCTACAT
OsPIP1-2R GTCTCGTACAGGCCCTTCTG
Os02g0629200 OsPIP2-2F CTGACCAAGTGGTCGCTGTA
OsPIP2-2R GTAGGACCCGAGCTTGTACC
Os04g0521100 OsPIP2-3F CGGTGTTCATGGTTCACTTG
OsPIP2-3R TGCCTTGTGCTGGTTGTAGA
Os07g0448100 OsPIP2-4F GCCGTGGTCTACAACAACAA
OsPIP2-4R GGATGACCTGGTGGTACAGC
Os03g0861300 OsPIP2-8F CACCGTGATCGGTGAGAAG
OsPIP2-8R GCAGTACACCAGCACGAAGA
Os03g0381200 β-Actin F AAGAAGGAGCAGCGCATTAC
β-Actin R CCTGATTGATCCCGACAAGT

图2

不同浓度HgCl2溶液下水稻叶片相对净光合速率和相对气孔导度随离体饲养时长的变化 C0~C4: HgCl2溶液浓度(0、100、200、300和500 µmol L-1); T0~T9: 离体饲养时长(0、0.5、1、1.5、2、2.5、3、3.5、4和4.5 h)。"

表2

不同HgCl2浓度下YLY 900和HLY 898 Fv/Fm随离体饲养时长的变化"

品种
Variety		 离体饲养时长
Feeding times
in vitro		 HgCl2浓度 HgCl2 concentration (µmol L-1)
C0 C1 C2 C3 C4
Y两优900
YLY900		 T0 0.840±0.002 0.839±0.003 0.840±0.002 0.837±0.003 0.839±0.003
T1 0.838±0.006 0.836±0.004 0.838±0.005 0.838±0.003 0.838±0.002
T2 0.837±0.004 0.836±0.004 0.836±0.004 0.834±0.010 0.838±0.002
T3 0.837±0.004 0.839±0.007 0.836±0.006 0.832±0.011 0.836±0.004
T4 0.839±0.001 0.836±0.001 0.837±0.003 0.834±0.007 0.836±0.003
T5 0.838±0.004 0.835±0.002 0.839±0.003 0.835±0.007 0.834±0.006
T6 0.837±0.004 0.836±0.002 0.838±0.002 0.837±0.002 0.838±0.003
T7 0.838±0.003 0.836±0.002 0.836±0.004 0.835±0.004 0.836±0.003
T8 0.833±0.005 0.833±0.002 0.838±0.002 0.836±0.004 0.836±0.003
T9 0.837±0.003 0.837±0.003 0.838±0.003 0.837±0.004 0.837±0.005
品种
Variety		 离体饲养时长
Feeding times
in vitro		 HgCl2浓度 HgCl2 concentration (µmol L-1)
C0 C1 C2 C3 C4
徽两优898
HLY898		 T0 0.840±0.007 0.839±0.002 0.841±0.005 0.842±0.005 0.842±0.002
T1 0.835±0.004 0.836±0.002 0.835±0.005 0.841±0.002 0.840±0.003
T2 0.834±0.002 0.833±0.002 0.835±0.004 0.839±0.001 0.833±0.016
T3 0.834±0.001 0.835±0.004 0.834±0.003 0.838±0.007 0.839±0.002
T4 0.835±0.003 0.840±0.004 0.834±0.010 0.842±0.002 0.839±0.003
T5 0.836±0.002 0.837±0.004 0.836±0.007 0.839±0.005 0.838±0.003
T6 0.837±0.003 0.836±0.004 0.839±0.004 0.841±0.002 0.838±0.005
T7 0.839±0.001 0.838±0.004 0.834±0.005 0.839±0.001 0.841±0.005
T8 0.837±0.004 0.839±0.003 0.836±0.003 0.838±0.004 0.840±0.003
T9 0.838±0.004 0.835±0.004 0.840±0.001 0.838±0.003 0.839±0.004

图3

两品种水稻相对气孔导度与相对净光合速率之间的相关性 浓度和时间处理同图2。"

图4

不同浓度HgCl2溶液下叶片相对含水量随离体处理时长的变化 浓度和时间处理同图2, 且柱上同一浓度不同小写字母表示在0.05水平上差异显著。"

图5

不同浓度HgCl2溶液下叶片SPAD值随离体饲养时长的变化 浓度和时间处理同图2, 且柱上同一浓度不同小写字母表示在0.05水平上差异显著。"

图6

不同浓度HgCl2溶液下YLY900水稻叶片表型随离体饲养时长(0 h和3 h)的变化 浓度处理同图2。"

表3

不同浓度HgCl2溶液下水稻SOD活性随离体饲养时长的变化"

HgCl2溶液浓度
Concentration of HgCl2 solutions (µmol L-1)		 SOD活性 SOD activity (U g-1)
T0 T2 T4 T6
C0 259.90±0.72 a 269.55±1.32 b 275.30±3.50 c 273.52±6.91 b
C1 259.50±0.76 a 273.91±2.43 ab 280.68±3.41 c 283.99±3.23 b
C2 259.78±2.51 a 278.86±3.07 a 322.16±2.95 a 292.84±5.24 a
C3 260.35±1.89 a 277.83±2.31 a 301.16±2.72 b 288.35±4.37 a
C4 260.11±0.89 a 255.03±4.05 c 242.87±2.85 d 229.94±2.49 c

表4

不同浓度HgCl2溶液下水稻POD活性随离体饲养时长的变化"

HgCl2溶液浓度
Concentration of HgCl2 solutions (µmol L-1)		 POD活性 POD activity (U g-1)
T0 T2 T4 T6
C0 165.52±2.67 a 173.14±1.35 e 179.22±1.46 e 182.65±1.19 e
C1 165.77±1.03 a 187.17±1.90 d 193.95±1.58 d 208.99±1.00 d
C2 166.00±0.23 a 199.40±0.79 c 227.29±0.70 c 237.39±1.96 c
C3 165.85±0.99 a 233.69±1.34 b 253.82±1.83 b 258.93±1.24 b
C4 165.52±0.63 a 264.12±2.31 a 271.80±0.62 a 278.88±1.31 a

表5

不同浓度HgCl2溶液下水稻MDA含量随离体饲养时长的变化"

HgCl2溶液浓度
Concentration of HgCl2 solutions (µmol L-1)		 MDA含量 MDA content (nmol g-1)
T0 T2 T4 T6
C0 15.64±0.34 a 17.01±0.25 d 18.36±0.45 e 20.74±0.43 c
C1 16.32±0.82 a 17.78±0.19 d 19.54±0.15 d 22.33±0.50 d
C2 16.29±0.32 a 25.80±0.20 c 28.22±0.67 c 31.27±0.26 c
C3 15.94±0.87 a 31.21±0.75 b 33.88±0.75 b 35.22±0.62 b
C4 16.08±0.53 a 42.28±0.64 a 45.19±0.56 a 47.40±0.43 a

图7

不同浓度HgCl2溶液下水稻质膜水通道蛋白基因相对表达随离体饲养时长的变化 浓度和时间处理同图2, 柱上同一离体饲养时长不同小写字母表示在0.05水平上差异显著。"

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