OsSAUR55 Regulates Rice Plant Type Through GA Pathway

doi:10.7668/hbnxb.20194611

Abstract

Abstract:

Knockout mutants ossaur55-1 and ossaur55-2 obtained by CRISPR/Cas9 technology in Francis background were used as research objects to study whether they affect the changes of agronomic traits such as plant type,leaf color,ear weight per plant and seed setting rate of rice.To explore whether OsSAUR55 regulates rice plant type by participating in GA pathway,and to provide theoretical basis for regulating rice plant type by related hormone pathway.The main agronomic traits of wild type(WT)and knockout mutant in adult plant stage were examined,including plant height,ear length,ear weight per plant,ten grain length,ten grain width,blade length,and blade width.Chlorophyll content and net photosynthetic rate of wild type and mutant leaf were measured by chlorophyll meter and photosynthetic rate meter.Real-time Fluorescence Quantitative PCR(qPCR)was used to determine the expression of OsSAUR55 gene in different tissues and different time points of rice seed germination.The subcellular localization of OsSAUR55 protein was observed by prokaryotic expression method.The contents of endogenous hormones(GA and IAA)in wild type and mutant were determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The expression of genes related to GA pathway was further analyzed.In the adult plant stage,mutant ossaur55-1 and ossaur55-2 showed significant phenotypic changes,in which the mutant was significantly shorter in plant height,greener in leaf color,increased in chlorophyll content,decreased in photosynthetic efficiency,decreased in seed setting rate,decreased in 1000-grain weight and decreased in panicle weight per plant than the wild type.The quantitative results showed that OsSAUR55 expression was the highest in the plumule,but not in the flag leaf.Confocal results showed that OsSAUR55 may be localized in cell membrane and nucleus.The results of hormone detection showed that the content of GA₄ was decreased significantly.Gene expression analysis showed that the expression levels of genes KAO,GA13ox1,GA20ox1 and KO1 in the GA synthesis pathway were significantly higher than those of the wild type,and the expression levels of GA2ox5,GA2ox8 and GA2ox9 in the GA metabolic pathway were also significantly higher than those of the wild type.It is preliminarily speculated that the deletion of OsSAUR55 gene led to the decrease of endogenous GA₄ content,which may regulate rice plant type through GA pathway,and then affect rice yield.

Key words: Rice, OsSAUR55, Plant type, CRISPR/Cas9, GA, Yield

PU Na, GAO Qiaoli, WANG Hui, LIU Jinzhao, CHEN Jialing, LUO Zengtong, XIAO Wuming, CHEN Chun. OsSAUR55 Regulates Rice Plant Type Through GA Pathway[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(2): 106-115. doi: 10.7668/hbnxb.20194611.

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Figures/Tables 10

References 56

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基因名称 Genes	引物序列(5'—3') The sequences of primers(5'—3')
β-Actin	F:TCTCTCAGCACATTCCAGCA	R:AATCACAAGTGAGAACCACAG
CPS1	F:TCAAGAGACACCGCCAGTTC	R:ACAGTGCATGACCCTGGATG
KS1	F:CTCGATCAGCTGCCATTTGC	R:GCGGGCTTCAGACAATTCAC
KAO	F:CTCCTTCGTGTCCTTCCGTC	R:ACCACAGCTGAACCTTCCAC
GA13ox1	F:TTCAACCCAGAGCGCTTCTC	R:CCATAGCAAAGCCCTGACCA
GA20ox1	F:TTCTTCCTCTGCCCGGAGAT	R:CATGTCGGCCCTGTAGTGG
GA3ox1	F:CGGCGATCTCTTCCATGTGC	R:ACCGAGGAAGTAGGGCATG
KO1	F:AACCCGCCTCCAGCTGTT	R:CAACTACGGAAGAAGCCCCG
GA2ox1	F:ACCCGCAGATCCTTAGCTTG	R:TGAACCCACATCTCCTTGCC
GA2ox3	F:ACTCGTTGCAGGTTCTGACC	R:GTGGCAATGGTGCAATCCTC
GA2ox5	F:GATGCCCATTACCGGAGCTT	R:AAGCGGTGCAGTCCAATCTT
GA2ox8	F:GAGCAGTGGAGTGCAGTTCA	R:CCAGCCGGTAAAAACCATGC
GA2ox9	F:AGCACAGGGTGATGACGAAC	R:CCTTGTAAGGGGAAGGCTCC
GA2ox10	F:TGAGTACGTTGTGGCGGTGA	R:AGACCAGCCTAACCAACGCA
qSAUR55	F:GGAGGAAGAGGCTTACAATGTC	R:AACACGTACTCCATCACCATG

基因名称 Genes	引物序列(5'—3') The sequences of primers(5'—3')
β-Actin	F:TCTCTCAGCACATTCCAGCA	R:AATCACAAGTGAGAACCACAG
CPS1	F:TCAAGAGACACCGCCAGTTC	R:ACAGTGCATGACCCTGGATG
KS1	F:CTCGATCAGCTGCCATTTGC	R:GCGGGCTTCAGACAATTCAC
KAO	F:CTCCTTCGTGTCCTTCCGTC	R:ACCACAGCTGAACCTTCCAC
GA13ox1	F:TTCAACCCAGAGCGCTTCTC	R:CCATAGCAAAGCCCTGACCA
GA20ox1	F:TTCTTCCTCTGCCCGGAGAT	R:CATGTCGGCCCTGTAGTGG
GA3ox1	F:CGGCGATCTCTTCCATGTGC	R:ACCGAGGAAGTAGGGCATG
KO1	F:AACCCGCCTCCAGCTGTT	R:CAACTACGGAAGAAGCCCCG
GA2ox1	F:ACCCGCAGATCCTTAGCTTG	R:TGAACCCACATCTCCTTGCC
GA2ox3	F:ACTCGTTGCAGGTTCTGACC	R:GTGGCAATGGTGCAATCCTC
GA2ox5	F:GATGCCCATTACCGGAGCTT	R:AAGCGGTGCAGTCCAATCTT
GA2ox8	F:GAGCAGTGGAGTGCAGTTCA	R:CCAGCCGGTAAAAACCATGC
GA2ox9	F:AGCACAGGGTGATGACGAAC	R:CCTTGTAAGGGGAAGGCTCC
GA2ox10	F:TGAGTACGTTGTGGCGGTGA	R:AGACCAGCCTAACCAACGCA
qSAUR55	F:GGAGGAAGAGGCTTACAATGTC	R:AACACGTACTCCATCACCATG

农艺性状 Agronomic character	WT	ossaur55-1	ossaur55-2
株高/cm Plant height	115.00±6.41a	85.90±12.70b	74.70±5.90b
剑叶长/cm Blade length	60.80±4.70a	30.30±5.46b	30.50±3.16b
剑叶宽/cm Blade width	1.55±0.05c	2.03±0.02a	1.92±0.10b
单株穗质量/g Ear weight per plant	13.36±0.94a	8.83±0.34b	8.53±1.04b
穗长/cm Panicle length	26.07±3.62a	20.84±1.10b	19.27±1.25b
单株穗数 Number of spikes per plant	5.00±1.00a	4.33±0.58a	4.67±0.58a
平均穗粒数 Average number of grains per spike	240.33±66.27a	194.33±42.59a	197.00±7.54a
结实率/% Setting rate	0.69±0.10a	0.68±0.04a	0.67±0.09a
千粒质量/g 1000-grain weight	19.02±0.25a	15.40±0.65c	16.89±0.37b
十粒长/cm Ten grains length	8.73±0.15a	8.77±0.25a	8.80±0.20a
十粒宽/cm Ten grains width	2.50±0.10a	2.50±0.10a	2.50±0.10a

农艺性状 Agronomic character	WT	ossaur55-1	ossaur55-2
株高/cm Plant height	115.00±6.41a	85.90±12.70b	74.70±5.90b
剑叶长/cm Blade length	60.80±4.70a	30.30±5.46b	30.50±3.16b
剑叶宽/cm Blade width	1.55±0.05c	2.03±0.02a	1.92±0.10b
单株穗质量/g Ear weight per plant	13.36±0.94a	8.83±0.34b	8.53±1.04b
穗长/cm Panicle length	26.07±3.62a	20.84±1.10b	19.27±1.25b
单株穗数 Number of spikes per plant	5.00±1.00a	4.33±0.58a	4.67±0.58a
平均穗粒数 Average number of grains per spike	240.33±66.27a	194.33±42.59a	197.00±7.54a
结实率/% Setting rate	0.69±0.10a	0.68±0.04a	0.67±0.09a
千粒质量/g 1000-grain weight	19.02±0.25a	15.40±0.65c	16.89±0.37b
十粒长/cm Ten grains length	8.73±0.15a	8.77±0.25a	8.80±0.20a
十粒宽/cm Ten grains width	2.50±0.10a	2.50±0.10a	2.50±0.10a

激素 Hormone	WT	ossaur55-1	ossaur55-2
IAA	5.54±0.04a	5.31±0.03a	5.50±0.10a
GA₁	2.09±0.03a	2.11±0.03a	2.13±0.02a
GA₃	0.22±0.01a	0.30±0.01a	0.26±0.01b
GA₄	0.87±0.01a	0.43±0.01b	0.36±0.01c
GA₇	未检出	未检出	未检出

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