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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (9): 2377-2389.doi: 10.3724/SP.J.1006.2022.12057

• TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

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 Online:2022-09-12 Published:2022-07-15
  • Contact: WU LI-Quan,HE Hai-Bing E-mail:1715994244@qq.com;hhb_agr@ahau.edu.cn
  • 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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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

Fig. 1

HgCl2 feeding treatment and photosynthetic gas exchange parameters of rice leaves in vitro"

Table 1

Name and primer sequence of aquaporin genes (OsPIPs)"

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

Fig. 2

Dynamic changes of relative net photosynthetic rate and relative stomatal conductance of rice leaves with times in different HgCl2 concentrations C0-C4: HgCl2 solution concentrations (0, 100, 200, 300, and 500 µmol L-1); T0-T9: feeding times in vitro (0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, and 4.5 h). YLY900: Y liangyou 900; HLY898: Huiliangyou 898."

Table 2

Dynamic changes of Fv/Fm of YLY 900 and HLY 898 with times in different HgCl2 concentrations"

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

Fig. 3

Correlation between relative stomatal conductance and relative net photosynthetic rate in two rice varieties The concentrations and time treatments are the same as those given in Fig. 2. YLY900: Y liangyou 900; HLY898: Huiliangyou 898."

Fig. 4

Dynamic changes of relative water content of leaves under different concentrations of HgCl2 solution with treatment times in vitro The concentration and time treatments are the same as those given in Fig. 2, and values followed by different lowercase letters above the bars at the same concentrations are significantly different at the 0.05 probability level."

Fig. 5

Dynamic changes of SPAD value of leaves under different concentrations of HgCl2 solution with treatment times in vitro The concentration and time treatments are the same as those given in Fig. 2, and values followed by different lowercase letters above the bars at the same concentrations are significantly different at the 0.05 probability level."

Fig. 6

Changes of leaf phenotype of YLY900 rice under different concentrations of HgCl2 solution with treatment times (0 h and 3 h) in vitro The concentration treatments are the same as those given in Fig. 2. YLY900: Y liangyou 900."

Table 3

Dynamic changes of SOD activity of rice leaves under different concentrations of HgCl2 solution with treatment times in vitro"

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

Table 4

Dynamic changes of POD activity of rice leaves under different concentrations of HgCl2 solution with treatment times in vitro"

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

Table 5

Dynamic changes of MDA content in rice under different concentrations of HgCl2 solution with treatment times in vitro"

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

Fig. 7

Relative expression of aquaporin gene in rice plasma membrane under different concentrations of HgCl2 solution with treatment times in vitro The concentrations and time treatments are the same as those given in Fig. 2, and values followed by different small letters above the bar at the same time of treatment in vitro are significantly different at the 0.05 probability level."

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