Biotechnology Bulletin ›› 2025, Vol. 41 ›› Issue (4): 211-226.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0766
SU Lin(
), QI Yu-cen(
), YANG Yi, CHENG He-feng, JI Si-yu, LIU Ting, CHENG Tang-ren, WANG Jia, ZHANG Qi-xiang, PAN Hui-tang(
)
Received:2024-08-08
Online:2025-04-26
Published:2025-04-25
Contact:
PAN Hui-tang
E-mail:sulin00125@163.com;htpan@bjfu.edu.cn
SU Lin, QI Yu-cen, YANG Yi, CHENG He-feng, JI Si-yu, LIU Ting, CHENG Tang-ren, WANG Jia, ZHANG Qi-xiang, PAN Hui-tang. Regulating the Resistance to Black Spot Disease by Xylanase Hydrolase Gene RcXYNC in Roses[J]. Biotechnology Bulletin, 2025, 41(4): 211-226.
| 序号 Serial number | 编号 Number | NCBI序列号 NCBI accession | 蛋白注释 Protein annotation |
|---|---|---|---|
| X1 | RC0G0005100 | XM_024324446.2 | 月季RNA结合蛋白MSI同源蛋白2 Rosa chinensis RNA-binding protein MSI homolog 2 |
| X2 | RC0G0013700 | XM_024326448.2 | 月季叶绿体a-b结合蛋白151 Rosa chinensis chlorophyll a-b binding protein 151 |
| X3 | RC0G0171100 | XM_024335925.2 | 月季60S核糖体蛋白L6 Rosa chinensis 60S ribosomal protein L6 |
| X4 | RC1G0312200 | XM_024317775.2 | 月季RING蛋白1a Rosa chinensis RING-box protein 1a |
| X5 | RC1G0321200 | XM_040516491.1 | 月季细胞分裂蛋白FtsY同源蛋白 Rosa chinensis cell division protein FtsY homolog |
| X6 | RC1G0417300 | XM_024334311.2 | 月季叶绿体a-b结合蛋白6A Rosa chinensis chlorophyll a-b binding protein 6A |
| X7 | RC1G0487500 | XM_024308438.1 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X8 | RC2G0046000 | XM_024329020.2 | 月季GTP结合核蛋白Ran-3 Rosa chinensis GTP-binding nuclear protein Ran-3 |
| X9 | RC2G0102900 | XM_040515202.1 | 月季20 kD分子伴侣蛋白 Rosa chinensis 20 kD chaperonin |
| X10 | RC2G0170900 | XM_024327498.2 | 月季放氧增强蛋白1 Rosa chinensis oxygen-evolving enhancer protein 1 |
| X11 | RC2G0252900 | XM_024330212.2 | 月季组蛋白H3.3 Rosa chinensis histone H3.3 |
| X12 | RC2G0273300 | XM_024330385.1 | 月季PLAT结构域蛋白3 Rosa chinensis PLAT domain-containing protein 3 |
| X13 | RC2G0334800 | XM_024326224.2 | 月季转录因子TGA2.3 Rosa chinensis transcription factor TGA2.3 |
| X14 | RC2G0337100 | XM_024329362.2 | 月季核酮糖二磷酸羧化酶小链 Rosa chinensis ribulose bisphosphate carboxylase small chain |
| X15 | RC2G0436200 | XM_024323609.2 | 月季磷脂酶 Rosa chinensis phospholipase D alpha 1 |
| X16 | RC2G0583300 | XM_024324653.2 | 月季蛋白翻译因子SUI1同源蛋白2 Rosa chinensis protein translation factor SUI1 homolog 2 |
| X17 | RC2G0682000 | XM_024324666.2 | 月季50S核糖体蛋白L27 Rosa chinensis 50S ribosomal protein L27 |
| X18 | RC3G0212600 | XR_002933950.2 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X19 | RC3G0287600 | XM_024331350.2 | 月季叶绿体a-b结合蛋白CP29.2 Rosa chinensis chlorophyll a-b binding protein CP29.2 |
| X20 | RC3G0321300 | XM_024335622.2 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X21 | RC4G0025300 | XM_024338987.2 | 月季3-酮酰基辅酶A合成酶11 Rosa chinensis 3-ketoacyl-CoA synthase 11 |
| X22 | RC4G0451500 | XM_024341654.2 | 月季泛素40S核糖体蛋白S27a Rosa chinensis ubiquitin-40S ribosomal protein S27a |
| X23 | RC4G0478900 | XM_024338228.2 | 月季转录因子MYB1R1 Rosa chinensis transcription factor MYB1R1 |
| X24 | RC5G0099800 | XM_024343555.2 | 月季14-3-3型D蛋白 Rosa chinensis 14-3-3-like protein D |
| X25 | RC5G0122400 | XM_024305557.2 | 月季叶绿体a-b结合蛋白LHCII 1型 Rosa chinensis chlorophyll a-b binding protein of LHCII type 1 |
| X26 | RC5G0122500 | XM_024302937.2 | 月季叶绿体a-b结合蛋白LHCII 1型 Rosa chinensis chlorophyll a-b binding protein of LHCII type 1 |
| X27 | RC5G0175700 | XM_024305213.2 | 月季光系统I反应中心亚基Ⅲ Rosa chinensis photosystem I reaction center subunit Ⅲ |
| X28 | RC5G0230100 | XM_024301146.2 | 月季翻译延长因子2 Rosa chinensis elongation factor 2 |
| X29 | RC5G0317100 | XM_024342352.2 | 月季叶绿体a-b结合蛋白3 Rosa chinensis chlorophyll a-b binding protein 3 |
| X30 | RC5G0380400 | XM_024301028.2 | 月季细胞分裂蛋白FtsZ同源蛋白1 Rosa chinensis cell division protein FtsZ homolog 1 |
| X31 | RC5G0648200 | XM_024342978.2 | 月季果糖二磷酸醛缩酶 Rosa chinensis fructose-bisphosphate aldolase |
| X32 | RC6G0082300 | XR_005801236.1 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X33 | RC6G0168800 | XM_024312327.2 | 月季26S蛋白酶体非ATPase调节亚基14同源蛋白 Rosa chinensis 26S proteasome non-ATPase regulatory subunit 14 homolog |
| X34 | RC6G0391500 | XM_024307241.2 | 月季光系统Ⅱ反应中心W蛋白 Rosa chinensis photosystem Ⅱ reaction center W protein |
| X35 | RC6G0394400 | XM_024305667.2 | 月季MPBQ转移酶 Rosa chinensis 2-methyl-6-phytyl-1, 4-hydroquinone methyltransferase |
| X36 | RC6G0457600 | XM_024306392.2 | 月季40S核糖体蛋白S11 Rosa chinensis 40S ribosomal protein S11 |
| X37 | RC6G0524600 | XM_024312257.2 | 月季硫氧还蛋白CDSP32 Rosa chinensis thioredoxin-like protein CDSP32 |
| X38 | RC7G0139800 | XM_024312740.2 | 月季28 kD核糖核蛋白 Rosa chinensis 28 kD ribonucleoprotein |
| X39 | RC7G0322800 | XM_024317397.2 | 月季磷酸核酮糖激酶 Rosa chinensis phosphoribulokinase |
| X40 | RC7G0482200 | XM_024312769.2 | 月季60S核糖体蛋白L13a-4 Rosa chinensis 60S ribosomal protein L13a-4 |
| X41 | RC7G0520700 | XM_024313678.2 | 月季表皮蜡质合成相关蛋白2 Rosa chinensis protein ECERIFERUM 2 |
Table 1 Candidate proteins potentially interacting with RcXYNC
| 序号 Serial number | 编号 Number | NCBI序列号 NCBI accession | 蛋白注释 Protein annotation |
|---|---|---|---|
| X1 | RC0G0005100 | XM_024324446.2 | 月季RNA结合蛋白MSI同源蛋白2 Rosa chinensis RNA-binding protein MSI homolog 2 |
| X2 | RC0G0013700 | XM_024326448.2 | 月季叶绿体a-b结合蛋白151 Rosa chinensis chlorophyll a-b binding protein 151 |
| X3 | RC0G0171100 | XM_024335925.2 | 月季60S核糖体蛋白L6 Rosa chinensis 60S ribosomal protein L6 |
| X4 | RC1G0312200 | XM_024317775.2 | 月季RING蛋白1a Rosa chinensis RING-box protein 1a |
| X5 | RC1G0321200 | XM_040516491.1 | 月季细胞分裂蛋白FtsY同源蛋白 Rosa chinensis cell division protein FtsY homolog |
| X6 | RC1G0417300 | XM_024334311.2 | 月季叶绿体a-b结合蛋白6A Rosa chinensis chlorophyll a-b binding protein 6A |
| X7 | RC1G0487500 | XM_024308438.1 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X8 | RC2G0046000 | XM_024329020.2 | 月季GTP结合核蛋白Ran-3 Rosa chinensis GTP-binding nuclear protein Ran-3 |
| X9 | RC2G0102900 | XM_040515202.1 | 月季20 kD分子伴侣蛋白 Rosa chinensis 20 kD chaperonin |
| X10 | RC2G0170900 | XM_024327498.2 | 月季放氧增强蛋白1 Rosa chinensis oxygen-evolving enhancer protein 1 |
| X11 | RC2G0252900 | XM_024330212.2 | 月季组蛋白H3.3 Rosa chinensis histone H3.3 |
| X12 | RC2G0273300 | XM_024330385.1 | 月季PLAT结构域蛋白3 Rosa chinensis PLAT domain-containing protein 3 |
| X13 | RC2G0334800 | XM_024326224.2 | 月季转录因子TGA2.3 Rosa chinensis transcription factor TGA2.3 |
| X14 | RC2G0337100 | XM_024329362.2 | 月季核酮糖二磷酸羧化酶小链 Rosa chinensis ribulose bisphosphate carboxylase small chain |
| X15 | RC2G0436200 | XM_024323609.2 | 月季磷脂酶 Rosa chinensis phospholipase D alpha 1 |
| X16 | RC2G0583300 | XM_024324653.2 | 月季蛋白翻译因子SUI1同源蛋白2 Rosa chinensis protein translation factor SUI1 homolog 2 |
| X17 | RC2G0682000 | XM_024324666.2 | 月季50S核糖体蛋白L27 Rosa chinensis 50S ribosomal protein L27 |
| X18 | RC3G0212600 | XR_002933950.2 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X19 | RC3G0287600 | XM_024331350.2 | 月季叶绿体a-b结合蛋白CP29.2 Rosa chinensis chlorophyll a-b binding protein CP29.2 |
| X20 | RC3G0321300 | XM_024335622.2 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X21 | RC4G0025300 | XM_024338987.2 | 月季3-酮酰基辅酶A合成酶11 Rosa chinensis 3-ketoacyl-CoA synthase 11 |
| X22 | RC4G0451500 | XM_024341654.2 | 月季泛素40S核糖体蛋白S27a Rosa chinensis ubiquitin-40S ribosomal protein S27a |
| X23 | RC4G0478900 | XM_024338228.2 | 月季转录因子MYB1R1 Rosa chinensis transcription factor MYB1R1 |
| X24 | RC5G0099800 | XM_024343555.2 | 月季14-3-3型D蛋白 Rosa chinensis 14-3-3-like protein D |
| X25 | RC5G0122400 | XM_024305557.2 | 月季叶绿体a-b结合蛋白LHCII 1型 Rosa chinensis chlorophyll a-b binding protein of LHCII type 1 |
| X26 | RC5G0122500 | XM_024302937.2 | 月季叶绿体a-b结合蛋白LHCII 1型 Rosa chinensis chlorophyll a-b binding protein of LHCII type 1 |
| X27 | RC5G0175700 | XM_024305213.2 | 月季光系统I反应中心亚基Ⅲ Rosa chinensis photosystem I reaction center subunit Ⅲ |
| X28 | RC5G0230100 | XM_024301146.2 | 月季翻译延长因子2 Rosa chinensis elongation factor 2 |
| X29 | RC5G0317100 | XM_024342352.2 | 月季叶绿体a-b结合蛋白3 Rosa chinensis chlorophyll a-b binding protein 3 |
| X30 | RC5G0380400 | XM_024301028.2 | 月季细胞分裂蛋白FtsZ同源蛋白1 Rosa chinensis cell division protein FtsZ homolog 1 |
| X31 | RC5G0648200 | XM_024342978.2 | 月季果糖二磷酸醛缩酶 Rosa chinensis fructose-bisphosphate aldolase |
| X32 | RC6G0082300 | XR_005801236.1 | 月季未知蛋白 Rosa chinensis uncharacterized |
| X33 | RC6G0168800 | XM_024312327.2 | 月季26S蛋白酶体非ATPase调节亚基14同源蛋白 Rosa chinensis 26S proteasome non-ATPase regulatory subunit 14 homolog |
| X34 | RC6G0391500 | XM_024307241.2 | 月季光系统Ⅱ反应中心W蛋白 Rosa chinensis photosystem Ⅱ reaction center W protein |
| X35 | RC6G0394400 | XM_024305667.2 | 月季MPBQ转移酶 Rosa chinensis 2-methyl-6-phytyl-1, 4-hydroquinone methyltransferase |
| X36 | RC6G0457600 | XM_024306392.2 | 月季40S核糖体蛋白S11 Rosa chinensis 40S ribosomal protein S11 |
| X37 | RC6G0524600 | XM_024312257.2 | 月季硫氧还蛋白CDSP32 Rosa chinensis thioredoxin-like protein CDSP32 |
| X38 | RC7G0139800 | XM_024312740.2 | 月季28 kD核糖核蛋白 Rosa chinensis 28 kD ribonucleoprotein |
| X39 | RC7G0322800 | XM_024317397.2 | 月季磷酸核酮糖激酶 Rosa chinensis phosphoribulokinase |
| X40 | RC7G0482200 | XM_024312769.2 | 月季60S核糖体蛋白L13a-4 Rosa chinensis 60S ribosomal protein L13a-4 |
| X41 | RC7G0520700 | XM_024313678.2 | 月季表皮蜡质合成相关蛋白2 Rosa chinensis protein ECERIFERUM 2 |
Fig. 1 Bioinformatics characteristics of RcXYNC geneA: Electrophoresis map of gene coding sequence cloning. B: Coding sequence of RcXYNC. C: Predicted conservative structure domain of RcXYNC. D: Multiple sequences alignment of RcXYNCwith other species. E: Phylogenetic analysis of RcXYNC (The location of RcXYNC marked with a '▲')
Fig. 2 Predicted secondary structure, tertiary structure, signal peptides and transmembrane domain of RcXYNCproteinA: Predicted secondary structure of RcXYNCprotein. B: Predicted tertiary structure of RcXYNCprotein. C: Predicted signal peptides of RcXYNC. D: Predicted transmembrane domain of RcXYNC
Fig. 4 Relative expression of RcXYNC in rose inoculated by H2O or M. rosaens: No significance; *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.000 1. The same below
Fig. 5 Analysis of RcXYNC promoter sequence and cis-acting elementsA: Electrophoresis map of gene promoter cloning. B: RcXYNC promoter sequence. C: Diagram of the cis-acting elements of the RcXYNC promoter
Fig. 6 Transient overexpression of RcXYNC enhanced the sensitiveness of rose to M. rosaeA: The black spot phenotype of R12-26 infected with M. rosae under RcXYNC overexpression on 8 d. B: Relative expression of RcXYNC of R12-26 infected with M. rosae under RcXYNC overexpression on 5 d. C: The black spot area statistic of R12-26 infected with M. rosae under RcXYNC overexpression on 8 d. D: The black spot phenotype of R13-54 infected with M. rosae under RcXYNC overexpression on 8 d. E: Relative expression of RcXYNC of R13-54 infected with M. rosae under RcXYNC overexpression on 5 d. F: The black spot area statistic of R13-54 infected with M. rosae under RcXYNC overexpression on 8 d. G: The H2O2 accumulation area statistic of R12-26 and R13-54 infected with M. rosae under RcXYNC overexpression on 2 d. H: The H2O2 accumulation phenotype of R12-26 and R13-54 infected with M. rosae under RcXYNC overexpression on 2 d. I: Relative expression of defense genes in R12-26 infected with M. rosae under RcXYNC overexpression on 2 d. J: Relative expression of defense genes in R13-54 infected with M. rosae under RcXYNC overexpression on 2 d (WT: rose leaves inoculated with M. rosae; PVX or PVX::RcXYNC; rose leaves infiltrated with Agrobacterium suspensions carrying PVX empty vector or PVX::RcXYNC recombinant vector and subsequently inoculated with M. rosae)
Fig. 7 Silencing of RcXYNC enhanced the resistance of rose to M. rosaeA: The black spot phenotype of R12-26 infected with M. rosae under RcXYNC silencing on 8 d. B: Relative expression of RcXYNC of R12-26 infected with M. rosae under RcXYNC silencing on 5 d. C: The black spot area statistic of R12-26 infected with M. rosae under RcXYNC silencing on 8 d. D: The black spot phenotype of R13-54 infected with M. rosae under RcXYNC silencing on 8 d. E: Relative expression of RcXYNC of R13-54 infected with M. rosae under RcXYNC silencing on 5 d. F: The black spot area statistic of R13-54 infected with M. rosae under RcXYNC silencing on 8 d. G: The H2O2 accumulation area statistic of R12-26 and R13-54 infected with M. rosae under RcXYNC silencing on 2 d. H: The H2O2 accumulation phenotype of R12-26 and R13-54 infected with M. rosae under RcXYNC silencing on 2 d. I: Relative expression of defense genes in R12-26 infected with M. rosae under RcXYNC silencing on 2 d. J: Relative expression of defense genes in R13-54 infected with M. rosae under RcXYNC silencing on 2 d (WT: rose leaves inoculated with M. rosae; TRV or TRV::RcXYNC; rose leaves infiltrated with Agrobacterium suspensions carrying TRV empty vector or TRV::RcXYNC recombinant vector and subsequently inoculated with M. rosae)
Fig. 8 Verifying the interaction between RcXYNC and candidate proteins via yeast two-hybridA: Self-activation identification of bait vector. B: Gene cloning of candidate proteins. (Lane M: DL2000 marker; other lanes: CDS cloning of candidate proteins) C: One-to-one validation of RcXYNC interaction with candidate target proteins in yeast (AD: pGADT7; BD: pGBKT7; AD-T+BD-Lam, AD+BD and AD+BD-RcXYNC: negative control; AD-T+BD-53: positive control; 10-1, 10-2, 10-3, 10-4: the dilution gradients of bacterial solution)
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