Light-responding Gene StRSM 1 Mediated by Chlorophyll-binding Protein CP24 Regulates Chlorophyll Accumulation

doi:10.13560/j.cnki.biotech.bull.1985.2020-0502

Abstract

Abstract:

Mutation on RSM（RADIALIS-like SANT/MYB）,a member of the single MYB transcription factor family inverts darkness-induced chlorophyll reduction,and the reason for it is unknown yet. To reveal how RSM regulates chlorophyll accumulation,StRSM 1-overexpressed positive transformant line of Nicotiana tabacum and Solanum tuberosum were generated,and chlorophyll accumulation and other physiologic phenotypes were assayed. The result showed that over-expression of RSM1 led to light green leaf and a less chlorophyll accumulation,and inhibition of RSM 1 expression increased chlorophyll accumulation. Expression assay of genes encoding chlorophyll metabolism and binding demonstrated that over-expression of StRSM 1 increased the expression of chlorophyll-binding protein CP24 under darkness and changed expression pattern of CP24. The above findings indicate that light-responding transcription factor StRSM 1 negatively regulates chlorophyll accumulation. CP24 involves in the regulation. The result will play an important role in further clarifying how RSM 1responds to light and deeply understanding the light response involved in RSM1.

Key words: StRSM 1, chlorophyll-binding protein, chlorophyll

SA Shi-juan, WU Han-yu, ZHANG Xiao-ping, ZHENG Rui, YAO Xin-ling. Light-responding Gene StRSM 1 Mediated by Chlorophyll-binding Protein CP24 Regulates Chlorophyll Accumulation[J]. Biotechnology Bulletin, 2021, 37(1): 198-204.

Figures/Tables 5

References 16

[1]	Katiyar A, Smita S, Lenka SK. et al. Genome-wide classification and expression analysis of MYB transcription factor families in rice and Arabidopsis[J]. BMC Genomics, 2012,13:544. doi: 10.1186/1471-2164-13-544 URL pmid: 23050870
[2]	Chen YH, Yang XY, He K, et al. The MYB transcription factor superfamily of Arabidopsis:expression analysis and phylogenetic comparison with the rice MYB family[J]. Plant Mol Biol, 2006,60:107-124. URL pmid: 16463103
[3]	Hamaguchi A, Yamashino T, Koizumi N, et al. A small subfamily of Arabidopsis RADIALIS-LIKE SANT/MYB genes:a link to HOOKLESS1-mediated signal transduction during early morphogenesis[J]. Biosci Biotechnol Biochem, 2008,72(10):2687-2696.
[4]	Park DY, Shim Y, Gi E, et al. The MYB-related transcription factor RADIALIS-LIKE3[OsRL3]functions in ABA-induced leaf senescence and salt sensitivity in rice[J]. Environ Exp Bot, 2018,156:86-95.
[5]	Mishanin VI, Trubitsin BV, Patsaeva SV, et al. Acclimation of shade-tolerant and light-resistant Tradescantia species to growth light:chlorophyll a fluorescence, electron transport, and xanthophyll content[J]. Photosynth Res, 2017,133(1-3):87-102. URL pmid: 28176042
[6]	Stolárik T, Hedtke B, Santrucek J, et al. Transcriptional and post-translational control of chlorophyll biosynjournal by dark-operative protochlorophyllide oxidoreductase in Norway spruce[J]. Photosynth Res, 2017,132(2):165-179. doi: 10.1007/s11120-017-0354-2 URL pmid: 28229362
[7]	Passarini F, Xu P, Caffarri S, et al. Towards in vivo mutation analysis:knock-out of specific chlorophylls bound to the light-harvesting complexes of Arabidopsis thaliana - the case of CP24(Lhcb6)[J]. Biochim Biophys Acta, 2014,1837(9):1500-1506. doi: 10.1016/j.bbabio.2014.02.012 URL pmid: 24561227
[8]	Ferroni L, Angeleri M, Pantaleoni L, et al. Light-dependent reversible phosphorylation of the minor photosystem II antenna Lhcb6(CP24)occurs in lycophytes[J]. Plant J, 2014,77(6):893-905.
[9]	Ruban VA. Light harvesting control in plant[J]. FEBS Letters, 2018,592:3030-3039. doi: 10.1002/1873-3468.13111 URL pmid: 29797317
[10]	Caffarri S, Kouril R, Kereïche S, et al. Functional architecture of higher plant photosystem II supercomplexes[J]. EMBO J, 2009,28:3052-3063. doi: 10.1038/emboj.2009.232 URL pmid: 19696744
[11]	Holleboom CP, Gacek DA, Liao PN. Carotenoid-chlorophyll coupling and fluorescence quenching in aggregated minor PSII proteins CP24 and CP29[J]. Photosynth Res, 2015,124(2):171-180.
[12]	Liu JF, Feng XJ, Cui XQ, et al. Expression of E. coli glgB in potatoes generating tuber starches with various features[J]. Molecular Plant Breeding, 2012,3(6):57-62.
[13]	Dekker JP, Boekema EJ. Supramolecular organization of thylakoid membrane proteins in green plants[J]. Biochim Biophys Acta, 2005,1706:12-39. doi: 10.1016/j.bbabio.2004.09.009 URL pmid: 15620363
[14]	Phillips MA, León P, Boronat A, et al. The plastidial MEP pathway:unified nomenclature and resources[J]. Trends in Plant Sci, 2008,13(12):619-623.
[15]	Cordoba E, Salmi M, León P. Unravelling the regulatory mechanisms that modulate the MEP pathway in higher plants[J]. J Exp Bot, 2009,60(10):2933.
[16]	Vranová E, Coman D, Gruissem W. Network analysis of the MVA and MEP pathways for isoprenoid synjournal[J]. Annu Rev Plant Biol, 2013,64:665-700. URL pmid: 23451776

引物名称	序列（5'-3'）	用途
RSMPF	acgagctcggtaccatgagttcgatg	StRSM 1体内过量表达载体构建
RSMPR	acggtaccgagctccttagtcagagg	StRSM 1体内过量表达载体构建
RSMMF	acggtaccggtaccatgagttcgatg	StRSM 1体内抑制表达载体构建
RSMMR	acgagctcgagctccttagtcagagg	StRSM 1体内抑制表达载体构建
qRSMF	acacggttcttcaggatcatgg	StRSM 1 mRNA积累定量测定
qRSMR	ttcttcagcagtcttgcctcca	StRSM 1 mRNA积累定量测定
SRSMF	cacggttcttcaggatcatg	重组载体阳性鉴定
SRSMR	ggggaagggcaccatac	重组载体阳性鉴定
L25F	cccctcaccacagagtctgc	StRSM 1体内表达定量测定内参
L25R	aagggtgttgttgtcctcaatctt	StRSM 1体内表达定量测定内参
612F	agaggaaaaatgcccaatatc	CP 24 mRNA积累的定性测定
612R	gcttatggggtttgtttgtg	CP 24 mRNA积累的定性测定

引物名称	序列（5'-3'）	用途
RSMPF	acgagctcggtaccatgagttcgatg	StRSM 1体内过量表达载体构建
RSMPR	acggtaccgagctccttagtcagagg	StRSM 1体内过量表达载体构建
RSMMF	acggtaccggtaccatgagttcgatg	StRSM 1体内抑制表达载体构建
RSMMR	acgagctcgagctccttagtcagagg	StRSM 1体内抑制表达载体构建
qRSMF	acacggttcttcaggatcatgg	StRSM 1 mRNA积累定量测定
qRSMR	ttcttcagcagtcttgcctcca	StRSM 1 mRNA积累定量测定
SRSMF	cacggttcttcaggatcatg	重组载体阳性鉴定
SRSMR	ggggaagggcaccatac	重组载体阳性鉴定
L25F	cccctcaccacagagtctgc	StRSM 1体内表达定量测定内参
L25R	aagggtgttgttgtcctcaatctt	StRSM 1体内表达定量测定内参
612F	agaggaaaaatgcccaatatc	CP 24 mRNA积累的定性测定
612R	gcttatggggtttgtttgtg	CP 24 mRNA积累的定性测定