OsDIS1 Negatively Regulates Rice Drought Tolerance Through Antioxidant Pathways

doi:10.13560/j.cnki.biotech.bull.1985.2022-0538

Abstract

Abstract:

Drought, as an abiotic stress factor, seriously affects the yield and quality of rice. Exploring drought-related functional genes will help to breed new drought-resistance varieties. In this study, the Nipponbare OsDIS1-RNAi transgenic rice was constructed by Agrobacterium-mediated transformation method, and a series of physiological and biochemical experiments and agronomic traits analysis were conducted on it and wild type. The results showed that the activities of antioxidant enzymes after drought treatment significantly enhanced, the content of malondialdehyde significantly reduced and the expression of reactive oxygen species clearance gene significantly increased in OsDIS1-RNAi transgenic rice compared with the wild type. After ABA treatment, OsDIS1-RNAi transgenic plants did not show significantly morphological changes; these results indicated that OsDIS1-mediated drought tolerance may be independent of the ABA pathway. Meanwhile, agronomic traits such as plant height, spike length and yield of OsDIS1-RNAi rice plants did not change. The above results indicate that OsDIS1 negatively regulates drought tolerance in rice by activating the antioxidant pathways to increase the intracellular scavenging capacity of reactive oxygen species, and it may have potential applications for drought tolerance breeding in rice.

Key words: rice, OsDIS1, antioxidant enzyme, drought tolerance, breeding

YANG Mao, LIN Yu-feng, DAI Yang-shuo, PAN Su-jun, PENG Wei-ye, YAN Ming-xiong, LI Wei, WANG Bing, DAI Liang-ying. OsDIS1 Negatively Regulates Rice Drought Tolerance Through Antioxidant Pathways[J]. Biotechnology Bulletin, 2023, 39(2): 88-95.

Figures/Tables 8

Table 1 Primers for RT-qPCR

引物Primer	序列Sequence（5'-3'）
Ubq-F	AACCAGCTGAGGCCCAAGA
Ubq-R	ACGATTGATTTAACCAGTCCATGA
OsDIS1-F	GTATCATCACAGTGCGAGAAT
OsDIS15-R DSM1-F DSM1-R	CATCTCATGAATTGCCATTAG GTGGTCATGGTCCATTATTGCC CCCAAACCCTCAACTGGCTTA
OsSIK1-F	TCTGCTAGTCTGCCCGAGGAA
OsSIK1-R	TATGTACTGGTTGCAATCAG
OsSKIPa-F	TACAGATGCGATCCAAGGTG
OsSKIPa-R	AGTGCCCTTAGCTCTTGCTC
OsDSM2-F	GCTTCGCCTGGCAAATGG
OsDSM2-R	ACCGTCCAAATGAGCTTCCA
OsBZIP23-F OsBZIP23-R	GGAGCAGCAAAAGAATGAGG GGTCTTCAGCTTCACCATCC

Fig. 1 Phenotypes of OsDIS1-RNAi transgenic plants treated with abscisic acid (ABA) A: OsDIS1-RNAi transgenic plants were germinated on 1/2 MS medium containing different concentrations of ABA. B: Seed germination rates of wild-type and OsDIS1-RNAi plants under different concentrations of ABA. C, D: OsDIS1-RNAi transgenic root length and plant height statistics of plants; RNAi-21 as the representative result of the graph, the same below. The error bar represents ± SD of triplicate experiments

Fig. 2 Expressions of ABA pathway genes in OsDIS1-RNAi plants detected by RT-qPCR

Fig. 3 Detection of reactive oxygen content of wild-type and RNAi

Fig. 4 Detection of antioxidant enzyme activity and MDA content in OsDIS1-RNAi transgenic plants A-C: CAT, SOD, POD activity and content in the leaves of OsDIS1-RNAi seedlings under drought stress. D: MDA content. The error bar represents ± SD of triplicate experiments

Fig. 5 Expressions of related reactive oxygen species （ROS）scavenging genes in OsDIS1-RNAi plants by RT-qPCR

Fig. 6 Growth phenotypes of OsDIS1-RNAi transgenic rice

Table 2 Agronomic characteristics of transgenic rice OsDIS1-RNAi

品种名称 Variety	株高 Plant height /cm	穗长 Panicle length /cm	单株有效穗数 Effective panicle per plant	每穗实粒数 Filled grains per panicle	每穗总粒数 Total grains per panicle	结实率 Seed setting /%	千粒重 1 000 grain weight/g	单株产量 Yield per plant /g
RNAi-8	94.4±5.2	19.4±1.6	26.8± 2.7	94.9±10.6	106.9±17.7	88.8±6.7	24.7±0.7	75.1±7.7
RNAi-21	96.7±6.2	20.4±1.4	28.6± 2.5	96.9±11.7	111.2±14.8	87.1±4.0	25.8±0.6	77.3±8.7
RNAi-22	95.7 ±4.1	19.6±0.9	30.4± 2.9	89.7±9.0	106.3±19.4	84.3±5.6	25.4±0.4	72.9±7.2
NPB	96.2±5.2	19.1±1.4	26.2±2.6	95.6±13.3	108.6±19.3	88.0±7.5	24.9±0.5	73.1±6.8

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