A proteomic approach to investigating the promotive effects of brassinolide on root growth of rice seedlings

doi:10.1007/s11703-011-1081-7

Front Agric Chin

2011, Vol. 5

Issue (2) : 146-151 https://doi.org/10.1007/s11703-011-1081-7

RESEARCH ARTICLE

A proteomic approach to investigating the promotive effects of brassinolide on root growth of rice seedlings

Shenghui LI, Fengru WANG(

), Jingao DONG(

)

Agricultural University of Hebei, Baoding 071000, China

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Abstract

Brassinolide influenced rice (Oryza sativa L.) root growth in a concentration-dependent manner. Roots grown in 0.1 nmol/L BL were 28% longer than those in the control. On the other hand, the roots grown in 1000 nmol/L BL were 65% shorter in comparison with those in the control. To investigate how BL influences rice root growth, proteome analysis techniques were applied. After BL treatments, total proteins from rice root were extracted separately. Extracted proteins were separated by two-dimensional polyacrylamide gel electrophoresis and analyzed using an automated protein sequencer and mass spectrometer. There were 33 proteins that showed differences in the accumulation levels as a result of treatments with BL. Proteins related to stress tolerance, enzymes, and cell structure were mainly found in the root. There were many proteins regulated by other hormone or light also. Results suggest that the physiologic functions of these proteins detected using powerful proteome analysis are implicated in root elongation triggered by BL. High-level brassinosteriod (BR) indicated that IAA amidohydrolase, which can release active IAA from IAA pool was increased, and the IAA level was so high that the root length was inhibited.

Keywords rice proteomics brassinolide root growth

Corresponding Author(s): WANG Fengru,Email:fengruwang@yahoo.com.cn; DONG Jingao,Email:shmdjg@hebau.edu.cn

Issue Date: 05 June 2011

Cite this article:

Shenghui LI,Fengru WANG,Jingao DONG. A proteomic approach to investigating the promotive effects of brassinolide on root growth of rice seedlings[J]. Front Agric Chin, 2011, 5(2): 146-151.

URL:

https://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1081-7
https://academic.hep.com.cn/fag/EN/Y2011/V5/I2/146

Tab.1 The root length of rice seedlings after a series of BL treatment for five days

Fig.1 The cell shape in rice root stained using PI and observed with confocal fluorescence microscope. A–C show the root top, representing control, 0.1 nmol/L BL treatment, and 1000 nmol/L BL treatment, respectively. D–F show the mature zone, representing control, 0.1 nmol/L BL treatment, and 1000 nmol/L BL treatment, respectively.

Spot no.	Code	Protein matched to	Conserved domain	Fold changes
0 nmol/L	0.1 nmol/L	1000 nmol/L
1	gi\|19920119	Putative retroelement	Reverse transcriptase	-1000000	1000000	-1000000
2	gi\|37535140	Mitochondrial chaperonin-60	TCP-1/cpn60 chaperonin family	1000000	-1000000	1000000
3	gi\|51535280	Putative PrMC3	Esterase_lipase	-1000000	1000000	-1000000
4	gi\|53749382	Putative ferredoxin sulfite reductase	Nitrite and sulphite reductase 4Fe-4S domain	6.86713	-6.86713	-2.67854
5	gi\|34894766	putative peroxidase	secretory peroxidases	4.55727	-4.55727	-1.99768
6	gi\|50910077	Translational elongation factor Tu	Elongation factor Tu GTP binding domain	12.0737	-12.0737	-1.78652
7	gi\|62733235	dnaK-type molecular chaperone hsp70- rice	Hsp70 protein	-4.97645	-10.7623	10.7623
8	gi\|1296955	osr40c1	Not putatively conserved domains	1000000	-1000000	-2.76531
9	gi\|50919967	Putative IAA amidohydrolase	Glutamate carboxypeptidases	-3,87622	-1000000	1000000
10	gi\|50918345	actin	actin	-1000000	1000000	-1000000
11	gi\|57900055	Putative ubiquitin-specific protease 6	Intracellular peptidases	-6.87645	-2.67543	6.87645
12	gi\|50931297	Unknown protein	Annexin repeats	-15.78462	15.78462	-1.76231
13	gi\|50923903	947OSJNBa0072K14.5	2-oxoacid dehydrogenases acyltransfer	-2.56435	-1.64310	2.56435
14	gi\|50924436	OSJNBa0006B20.1	Regulator of chromosome condensation	1000000	-1000000	-1000000
15	gi\|50933679	Putative peroxidase 1 precursor	Peroxidase	1000000	-1000000	-3.87465
16	gi\|50946591	Putative activator of 90 kDa heat shock protein ATPase homolog 1	Heat shock protein	-20.23759	20.23759	-20.23759
17	gi\|53749369	Putative 1,4-benzoquinone reductase	Flavodoxin_1	1000000	-1000000	-1000000
18	gi\|34908134	Osmotin-like protein	Thaumatin family	-1.87492	-30.64921	30.64921
19	gi\|56784135	Putative Y1 protein	WD40 domain (signal transduction)	-2.98641	1000000	-1000000
20	gi\|295885	Actin	Actin	-1.58299	3.652491	-3.652491
21	gi\|303844	Eukaryotic initiation factor 4A	DEAD-box helicases, helicase superfamily c-terminal domain	-1000000	1000000	-1000000
22	gi\|50918875	Putative UDPglucose dehydrogenase	UDPglucose/GDP-mannose dehydrogenase family	20.38299	-1.99869	-20.38299
23	gi\|2662343	EF-1 alpha	Elongation factor Tu GTP binding domain	1000000	-1000000	1000000
24	gi\|50940457	Putative fructokinase	Fructokinases	-1000000	-1000000	1000000
25	gi\|50898962	Putative D-protein	Kelch motif	2.05292	-2.05292	-0.67239
26	gi\|50935067	Putative stationary phase survival protein SurE	Survival protein SurE	1000000	-1000000	1000000
27	gi\|49532749	Radc1	Not putatively conserved domains	-0.63927	2.21973	-2.21973
28	gi\|46798901	Kelch repeat containing protein	Kelch motif	-1000000	1000000	-1000000
29	gi\|50918681	Cysteine synthase	Pyridoxal-phosphate dependent enzyme	2.7625	3.9254	-2.7625
30	gi\|50355737	Putative DnaJ like protein	DnaJ domains	-3.01862	-1.09217	3.01862
31	gi\|50918709	Putative exoglucanase precursor	Glycosyl hydrolase family 3 domain..	-2.15790	2.15790	-2.08721
32	gi\|34897872	Putative phosphogluconate dehydrogenase	6-phosphogluconate dehydrogenase	-1000000	-1000000	1000000
33	gi\|34895730	Unknown protein	Mitochondrial glycoprotein	-3.98092	-4.53826	4.53826

Tab.2 Proteins regulated by BR in root total protein after 6 h BL treatment

Fig.2 IAA amidohydrolase (part of 2D-PAGE of root protein). A–C represent control, 0.1 nmol/L BL treatment, and 1000 nmol/L BL treatment, respectively.

Fig.3 RT-PCR of IAA amidohydrolase. The left is control, the middle is 0.1 nmol/L BL, and the right is 1000 nmol/L BL.

Tab.3 The IAA content of rice root after different concentration BL treatment for six hours

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