浙江农业学报 ›› 2022, Vol. 34 ›› Issue (2): 207-220.DOI: 10.3969/j.issn.1004-1524.2022.02.01
收稿日期:2021-05-21
出版日期:2022-02-25
发布日期:2022-03-02
作者简介:张斌,E-mail: zhangbin27104@163.com通讯作者:
张斌
基金资助:
YANG Xinxiaa(
), TANG Manshengb, ZHANG Binb,*(
)
Received:2021-05-21
Online:2022-02-25
Published:2022-03-02
Contact:
ZHANG Bin
摘要:
植物蛋白磷酸酶2C(PP2C)能够通过影响植物体内多种生物学过程在环境胁迫响应中发挥重要作用。为研究栽培大豆PP2C家族成员在盐胁迫中的功能,通过生物信息学手段结合转录组学分析对栽培大豆PP2C家族进行了系统探究。在栽培大豆中共鉴定出126个PP2C家族成员,并将其在进化树上分为11个亚家族,同一亚家族的成员具有类似的基因和蛋白结构。盐胁迫转录组测序结果显示,大豆126个GmPP2C基因中有9个是差异表达基因,其中,GmPP2C41、GmPP2C48、GmPP2C72、GmPP2C84、GmPP2C89、GmPP2C107和GmPP2C113在盐处理后表达水平上调,GmPP2C27和GmPP2C114在盐处理后表达下调。qRT-PCR结果显示,GmPP2C89和GmPP2C113在盐处理6 h表达水平上调最高。GO富集分析发现,9个差异基因主要具有离子结合和磷酸酶活性;Y2H结果显示,GmPP2C113和GmPP2C47能够互作。以上结果将为进一步研究大豆PP2C家族成员在盐胁迫响应中的功能与分子机制提供理论依据,为大豆耐盐性遗传改良提供重要候选基因。
中图分类号:
杨昕霞, 唐满生, 张斌. 大豆PP2C家族基因鉴定与响应盐胁迫的转录组分析[J]. 浙江农业学报, 2022, 34(2): 207-220.
YANG Xinxia, TANG Mansheng, ZHANG Bin. Identification of soybean PP2C family genes and transcriptome analysis in response to salt stress[J]. Acta Agriculturae Zhejiangensis, 2022, 34(2): 207-220.
| 基因名 Gene name | 正向引物 Forward primer(5'-3') | 反向引物 Reverse primer(5'-3') | 片段长度 Fragment length/bp |
|---|---|---|---|
| GmPP2C27 | AGGCTGCAGCAACCCATATT | GGCACAGAAGTAGGTGGCTT | 115 |
| GmPP2C41 | CCGCTTCATTTCTTCGGCG | CTTCTCTGCCATTCAGCGGC | 134 |
| GmPP2C48 | GTCTATGATGGGCATGGGGG | CCGTCCCTCCCATTCTTTCC | 122 |
| GmPP2C72 | GCCAAAGTGGAAATCCACGAC | CCATGTCTCTTCTCCTGCCG | 124 |
| GmPP2C84 | CCGAGACTTCTTCTCCCATCG | GAGAAGCGCGCCTATCCAG | 150 |
| GmPP2C89 | TCTTCCCTGCGAGTGGAGTC | GGATCTTCGTCTGGCAAGTGT | 112 |
| GmPP2C107 | ACTGGACACAAGGTGACGAA | TCACGCCGAACTTGGGAGAT | 118 |
| GmPP2C113 | ACCAGAAGTGACAGTGACCC | GATTCCCAACTCCATCGCAC | 158 |
| GmPP2C114 | GATCACCTCTTCCAGCACCT | ACACCAACCAGACAACAGGA | 173 |
表1 qRT-PCR引物
Table 1 Primers used for qRT-PCR
| 基因名 Gene name | 正向引物 Forward primer(5'-3') | 反向引物 Reverse primer(5'-3') | 片段长度 Fragment length/bp |
|---|---|---|---|
| GmPP2C27 | AGGCTGCAGCAACCCATATT | GGCACAGAAGTAGGTGGCTT | 115 |
| GmPP2C41 | CCGCTTCATTTCTTCGGCG | CTTCTCTGCCATTCAGCGGC | 134 |
| GmPP2C48 | GTCTATGATGGGCATGGGGG | CCGTCCCTCCCATTCTTTCC | 122 |
| GmPP2C72 | GCCAAAGTGGAAATCCACGAC | CCATGTCTCTTCTCCTGCCG | 124 |
| GmPP2C84 | CCGAGACTTCTTCTCCCATCG | GAGAAGCGCGCCTATCCAG | 150 |
| GmPP2C89 | TCTTCCCTGCGAGTGGAGTC | GGATCTTCGTCTGGCAAGTGT | 112 |
| GmPP2C107 | ACTGGACACAAGGTGACGAA | TCACGCCGAACTTGGGAGAT | 118 |
| GmPP2C113 | ACCAGAAGTGACAGTGACCC | GATTCCCAACTCCATCGCAC | 158 |
| GmPP2C114 | GATCACCTCTTCCAGCACCT | ACACCAACCAGACAACAGGA | 173 |
| 基因名 Gene name | 基因登录号 Gene ID | 外显子数量 Exon count | 氨基酸残基 Peptide residues | 分子量 Molecular weight/ku | 等电点 pI | 不稳定系数 Instability index | 亲水性 Grand average of hydropathicity | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| GmPP2C1 | GLYMA.01G127600 | 4 | 290 | 32 192.77 | 6.65 | 37.11 | -0.267 | ||||
| GmPP2C2 | GLYMA.01G158000 | 4 | 374 | 40 365.95 | 5.42 | 55.69 | -0.200 | ||||
| GmPP2C3 | GLYMA.01G191300 | 4 | 377 | 41 796.69 | 6.43 | 40.61 | -0.200 | ||||
| GmPP2C4 | GLYMA.01G199600 | 4 | 753 | 83 208.77 | 5.21 | 50.37 | -0.575 | ||||
| GmPP2C5 | GLYMA.01G225100 | 3 | 324 | 35 093.05 | 6.75 | 47.55 | -0.228 | ||||
| GmPP2C6 | GLYMA.02G009100 | 4 | 396 | 43 461.91 | 4.78 | 51.36 | -0.213 | ||||
| GmPP2C7 | GLYMA.02G044700 | 5 | 394 | 43 795.09 | 8.79 | 47.28 | -0.250 | ||||
| GmPP2C8 | GLYMA.02G058600 | 4 | 364 | 39 141.87 | 5.69 | 34.82 | -0.152 | ||||
| GmPP2C9 | GLYMA.02G144500 | 22 | 1 073 | 121 060.70 | 5.77 | 38.93 | -0.383 | ||||
| GmPP2C10 | GLYMA.02G228200 | 4 | 389 | 41 812.92 | 6.28 | 55.94 | -0.294 | ||||
| GmPP2C11 | GLYMA.02G250200 | 4 | 407 | 44 933.13 | 6.67 | 53.82 | -0.361 | ||||
| GmPP2C12 | GLYMA.03G043100 | 5 | 211 | 23 690.01 | 7.67 | 37.86 | -0.332 | ||||
| GmPP2C13 | GLYMA.03G044100 | 3 | 222 | 24 615.08 | 6.43 | 40.96 | -0.285 | ||||
| GmPP2C14 | GLYMA.03G176100 | 5 | 357 | 39 901.23 | 6.84 | 41.18 | -0.287 | ||||
| GmPP2C15 | GLYMA.03G225800 | 4 | 887 | 99 448.89 | 6.01 | 49.22 | -0.544 | ||||
| GmPP2C16 | GLYMA.03G233300 | 8 | 426 | 45 561.23 | 7.98 | 38.38 | -0.074 | ||||
| GmPP2C17 | GLYMA.03G233500 | 8 | 371 | 41 097.80 | 6.64 | 39.60 | -0.246 | ||||
| GmPP2C18 | GLYMA.04G038000 | 11 | 429 | 46 556.83 | 5.28 | 41.90 | -0.288 | ||||
| GmPP2C19 | GLYMA.04G053800 | 6 | 544 | 59 216.90 | 4.86 | 43.36 | -0.216 | ||||
| GmPP2C20 | GLYMA.04G059000 | 9 | 312 | 33 540.32 | 5.02 | 37.01 | -0.258 | ||||
| GmPP2C21 | GLYMA.04G069900 | 4 | 370 | 40 592.64 | 5.29 | 58.10 | -0.389 | ||||
| GmPP2C22 | GLYMA.04G101400 | 6 | 301 | 33 228.71 | 7.70 | 41.29 | -0.313 | ||||
| GmPP2C23 | GLYMA.05G113400 | 4 | 696 | 76 424.07 | 5.57 | 45.86 | -0.371 | ||||
| GmPP2C24 | GLYMA.05G116800 | 6 | 282 | 30 673.49 | 6.66 | 38.30 | -0.357 | ||||
| GmPP2C25 | GLYMA.05G227100 | 4 | 384 | 42 155.30 | 5.36 | 67.38 | -0.348 | ||||
| GmPP2C26 | GLYMA.06G016100 | 4 | 385 | 41 908.80 | 5.28 | 52.86 | -0.179 | ||||
| GmPP2C27 | GLYMA.06G039000 | 9 | 429 | 46 694.27 | 5.29 | 41.02 | -0.230 | ||||
| GmPP2C28 | GLYMA.06G050900 | 5 | 343 | 37 342.22 | 5.35 | 33.80 | -0.234 | ||||
| GmPP2C29 | GLYMA.06G054000 | 6 | 544 | 58 934.53 | 4.83 | 45.69 | -0.182 | ||||
| GmPP2C30 | GLYMA.06G059700 | 9 | 314 | 33 766.48 | 5.18 | 40.20 | -0.327 | ||||
| GmPP2C31 | GLYMA.06G071900 | 5 | 370 | 40 595.79 | 5.30 | 56.73 | -0.365 | ||||
| GmPP2C32 | GLYMA.06G103100 | 6 | 293 | 32 415.53 | 7.09 | 41.39 | -0.432 | ||||
| GmPP2C33 | GLYMA.06G130900 | 11 | 392 | 43 150.91 | 5.84 | 50.68 | -0.155 | ||||
| GmPP2C34 | GLYMA.06G238200 | 5 | 282 | 30 849.97 | 5.87 | 41.78 | -0.313 | ||||
| GmPP2C35 | GLYMA.06G296200 | 5 | 471 | 51 859.97 | 5.50 | 43.70 | -0.504 | ||||
| GmPP2C36 | GLYMA.07G131800 | 4 | 602 | 66 123.01 | 5.13 | 38.86 | -0.359 | ||||
| GmPP2C37 | GLYMA.07G233000 | 2 | 386 | 42 549.56 | 5.04 | 61.31 | -0.056 | ||||
| GmPP2C38 | GLYMA.07G239000 | 5 | 374 | 41 517.38 | 7.81 | 35.89 | -0.192 | ||||
| GmPP2C39 | GLYMA.07G244700 | 7 | 367 | 41 031.05 | 8.29 | 30.04 | -0.273 | ||||
| GmPP2C40 | GLYMA.07G248000 | 5 | 520 | 56 834.23 | 4.99 | 41.85 | -0.192 | ||||
| GmPP2C41 | GLYMA.08G033800 | 4 | 385 | 42 285.55 | 5.26 | 63.59 | -0.343 | ||||
| GmPP2C42 | GLYMA.08G071900 | 7 | 282 | 30 694.42 | 6.17 | 42.66 | -0.362 | ||||
| GmPP2C43 | GLYMA.08G081400 | 8 | 351 | 39 455.77 | 4.86 | 43.14 | -0.695 | ||||
| GmPP2C44 | GLYMA.08G178800 | 5 | 280 | 30 455.45 | 6.25 | 34.28 | -0.323 | ||||
| GmPP2C45 | GLYMA.09G031700 | 2 | 405 | 44 584.63 | 5.01 | 61.41 | -0.104 | ||||
| GmPP2C46 | GLYMA.09G034800 | 4 | 374 | 41 362.15 | 7.25 | 44.03 | -0.129 | ||||
| GmPP2C47 | GLYMA.09G045100 | 6 | 409 | 45 167.96 | 4.84 | 38.69 | -0.224 | ||||
| GmPP2C48 | GLYMA.09G066500 | 5 | 522 | 57 118.04 | 5.42 | 42.20 | -0.545 | ||||
| GmPP2C49 | GLYMA.09G096600 | 4 | 381 | 42 730.21 | 5.04 | 44.72 | -0.384 | ||||
| GmPP2C50 | GLYMA.09G111300 | 5 | 385 | 43 080.32 | 8.71 | 46.63 | -0.205 | ||||
| GmPP2C51 | GLYMA.09G249700 | 5 | 489 | 54 209.69 | 5.53 | 46.34 | -0.556 | ||||
| GmPP2C52 | GLYMA.09G280700 | 5 | 434 | 48 642.41 | 5.11 | 42.79 | -0.504 | ||||
| GmPP2C53 | GLYMA.10G004200 | 5 | 282 | 32 656.61 | 9.57 | 47.97 | -0.498 | ||||
| GmPP2C54 | GLYMA.10G009800 | 5 | 396 | 43 572.14 | 4.76 | 49.32 | -0.154 | ||||
| GmPP2C55 | GLYMA.10G047500 | 5 | 371 | 40 953.34 | 6.43 | 42.05 | -0.316 | ||||
| GmPP2C56 | GLYMA.10G149600 | 8 | 428 | 45 996.65 | 7.49 | 37.66 | -0.149 | ||||
| GmPP2C57 | GLYMA.10G150100 | 7 | 368 | 41 126.43 | 5.78 | 39.61 | -0.360 | ||||
| GmPP2C58 | GLYMA.10G182400 | 6 | 277 | 31 868.60 | 9.17 | 54.09 | -0.455 | ||||
| GmPP2C59 | GLYMA.10G259400 | 4 | 378 | 42 452.47 | 8.13 | 42.60 | -0.233 | ||||
| GmPP2C60 | GLYMA.10G271500 | 11 | 431 | 46 614.22 | 7.99 | 38.89 | -0.240 | ||||
| GmPP2C61 | GLYMA.10G282900 | 5 | 397 | 43 516.33 | 7.28 | 51.12 | -0.222 | ||||
| GmPP2C62 | GLYMA.10G291800 | 9 | 320 | 34 490.27 | 5.02 | 41.37 | -0.275 | ||||
| GmPP2C63 | GLYMA.10G294400 | 5 | 389 | 43 106.25 | 8.10 | 46.29 | -0.246 | ||||
| GmPP2C64 | GLYMA.10G297900 | 2 | 208 | 22 659.85 | 5.57 | 32.40 | 0.014 | ||||
| GmPP2C65 | GLYMA.11G018000 | 4 | 336 | 36 484.61 | 7.89 | 45.57 | -0.240 | ||||
| GmPP2C66 | GLYMA.11G042300 | 4 | 739 | 82 047.64 | 5.28 | 53.20 | -0.576 | ||||
| GmPP2C67 | GLYMA.11G050900 | 4 | 375 | 41 826.79 | 5.83 | 40.09 | -0.194 | ||||
| GmPP2C68 | GLYMA.11G087000 | 4 | 374 | 40 313.84 | 5.51 | 56.60 | -0.100 | ||||
| GmPP2C69 | GLYMA.11G139500 | 4 | 720 | 79 617.51 | 5.53 | 36.87 | -0.449 | ||||
| GmPP2C70 | GLYMA.11G170200 | 4 | 335 | 36 549.43 | 6.71 | 51.93 | -0.293 | ||||
| GmPP2C71 | GLYMA.11G171600 | 4 | 336 | 36 663.53 | 6.71 | 51.81 | -0.302 | ||||
| GmPP2C72 | GLYMA.11G222600 | 4 | 401 | 43 834.33 | 5.51 | 56.39 | -0.438 | ||||
| GmPP2C73 | GLYMA.12G063100 | 4 | 720 | 79 940.84 | 5.37 | 40.75 | -0.482 | ||||
| GmPP2C74 | GLYMA.12G109500 | 5 | 471 | 51 711.72 | 5.30 | 42.12 | -0.494 | ||||
| GmPP2C75 | GLYMA.12G116800 | 5 | 281 | 31 172.50 | 6.07 | 39.18 | -0.359 | ||||
| GmPP2C76 | GLYMA.12G202400 | 7 | 474 | 51 766.93 | 5.04 | 45.62 | -0.392 | ||||
| GmPP2C77 | GLYMA.12G226100 | 5 | 265 | 29 350.15 | 6.51 | 53.65 | -0.459 | ||||
| GmPP2C78 | GLYMA.13G042100 | 11 | 356 | 39 260.01 | 5.17 | 42.38 | -0.346 | ||||
| GmPP2C79 | GLYMA.13G106800 | 6 | 536 | 58 563.88 | 5.07 | 47.95 | -0.386 | ||||
| GmPP2C80 | GLYMA.13G135300 | 5 | 371 | 40 947.29 | 6.47 | 40.61 | -0.342 | ||||
| GmPP2C81 | GLYMA.13G165300 | 4 | 383 | 42 454.17 | 5.18 | 49.88 | -0.364 | ||||
| GmPP2C82 | GLYMA.13G274800 | 5 | 283 | 31 107.25 | 7.70 | 47.17 | -0.433 | ||||
| GmPP2C83 | GLYMA.13G299300 | 5 | 475 | 52 153.36 | 5.31 | 46.32 | -0.446 | ||||
| GmPP2C84 | GLYMA.14G066400 | 4 | 400 | 44 150.19 | 6.05 | 60.37 | -0.345 | ||||
| GmPP2C85 | GLYMA.14G082200 | 11 | 428 | 46 492.90 | 5.29 | 42.01 | -0.276 | ||||
| GmPP2C86 | GLYMA.14G104900 | 9 | 338 | 36 587.85 | 5.15 | 44.51 | -0.193 | ||||
| GmPP2C87 | GLYMA.14G108400 | 5 | 557 | 60 815.60 | 4.62 | 41.13 | -0.205 | ||||
| GmPP2C88 | GLYMA.14G161400 | 10 | 356 | 39 293.99 | 5.47 | 41.18 | -0.426 | ||||
| GmPP2C89 | GLYMA.14G162100 | 3 | 386 | 42 074.51 | 6.73 | 46.77 | -0.333 | ||||
| GmPP2C90 | GLYMA.14G195200 | 4 | 390 | 41 811.95 | 6.11 | 55.12 | -0.261 | ||||
| GmPP2C91 | GLYMA.15G053700 | 5 | 278 | 30 487.53 | 6.31 | 35.14 | -0.282 | ||||
| GmPP2C92 | GLYMA.15G139500 | 4 | 372 | 41 084.92 | 7.69 | 46.14 | -0.123 | ||||
| GmPP2C93 | GLYMA.15G172500 | 8 | 527 | 57 849.70 | 5.22 | 44.75 | -0.541 | ||||
| GmPP2C94 | GLYMA.15G201300 | 4 | 379 | 42 328.00 | 5.46 | 45.59 | -0.353 | ||||
| GmPP2C95 | GLYMA.16G122200 | 6 | 394 | 43 876.13 | 8.79 | 47.20 | -0.263 | ||||
| GmPP2C96 | GLYMA.17G026100 | 5 | 524 | 57 138.86 | 5.52 | 37.84 | -0.167 | ||||
| GmPP2C97 | GLYMA.17G029200 | 7 | 376 | 41 908.18 | 7.99 | 29.58 | -0.233 | ||||
| GmPP2C98 | GLYMA.17G034500 | 4 | 375 | 41 437.23 | 7.80 | 33.16 | -0.208 | ||||
| GmPP2C99 | GLYMA.17G038000 | 2 | 380 | 41 708.68 | 5.09 | 63.88 | -0.022 | ||||
| GmPP2C100 | GLYMA.17G052500 | 7 | 538 | 58 548.81 | 4.97 | 48.37 | -0.374 | ||||
| GmPP2C101 | GLYMA.17G105800 | 4 | 383 | 42 290.77 | 5.11 | 47.42 | -0.344 | ||||
| GmPP2C102 | GLYMA.17G153800 | 5 | 701 | 77 629.50 | 5.50 | 49.90 | -0.447 | ||||
| GmPP2C103 | GLYMA.17G218700 | 8 | 558 | 60 792.54 | 4.62 | 38.16 | -0.208 | ||||
| GmPP2C104 | GLYMA.17G221100 | 9 | 338 | 36 655.97 | 5.09 | 41.29 | -0.188 | ||||
| GmPP2C105 | GLYMA.17G230600 | 5 | 344 | 37 627.48 | 5.07 | 31.99 | -0.240 | ||||
| GmPP2C106 | GLYMA.17G243100 | 12 | 428 | 46 575.99 | 5.23 | 45.89 | -0.294 | ||||
| GmPP2C107 | GLYMA.18G035000 | 4 | 400 | 44 067.66 | 5.17 | 58.50 | -0.443 | ||||
| GmPP2C108 | GLYMA.18G061000 | 4 | 347 | 37 640.64 | 5.81 | 51.88 | -0.222 | ||||
| GmPP2C109 | GLYMA.18G180400 | 4 | 597 | 65 974.13 | 5.37 | 37.54 | -0.349 | ||||
| GmPP2C110 | GLYMA.18G208100 | 5 | 424 | 47 458.29 | 5.24 | 45.39 | -0.473 | ||||
| GmPP2C111 | GLYMA.18G242700 | 6 | 487 | 54 026.40 | 6.10 | 42.66 | -0.578 | ||||
| GmPP2C112 | GLYMA.18G283300 | 6 | 455 | 50 115.64 | 5.30 | 37.19 | -0.325 | ||||
| GmPP2C113 | GLYMA.19G069200 | 3 | 377 | 40 769.73 | 5.68 | 51.67 | -0.354 | ||||
| GmPP2C114 | GLYMA.19G147800 | 4 | 391 | 44 150.56 | 7.69 | 53.67 | -0.278 | ||||
| GmPP2C115 | GLYMA.19G176600 | 5 | 370 | 41 129.64 | 8.22 | 40.69 | -0.272 | ||||
| GmPP2C116 | GLYMA.19G222700 | 4 | 887 | 99 137.72 | 6.01 | 49.65 | -0.516 | ||||
| GmPP2C117 | GLYMA.19G230200 | 8 | 429 | 46 091.00 | 7.97 | 36.52 | -0.092 | ||||
| GmPP2C118 | GLYMA.19G230600 | 9 | 369 | 40 812.38 | 6.09 | 35.91 | -0.243 | ||||
| GmPP2C119 | GLYMA.20G106800 | 6 | 397 | 43 657.42 | 7.28 | 50.98 | -0.270 | ||||
| GmPP2C120 | GLYMA.20G119100 | 12 | 431 | 46 513.07 | 7.56 | 38.39 | -0.235 | ||||
| GmPP2C121 | GLYMA.20G131500 | 4 | 373 | 41 357.11 | 6.93 | 40.87 | -0.180 | ||||
| GmPP2C122 | GLYMA.20G207600 | 6 | 277 | 31 730.49 | 9.44 | 51.96 | -0.467 | ||||
| GmPP2C123 | GLYMA.20G238100 | 8 | 367 | 41 147.49 | 6.14 | 41.32 | -0.389 | ||||
| GmPP2C124 | GLYMA.20G238700 | 8 | 428 | 46 223.91 | 6.91 | 37.92 | -0.181 | ||||
| GmPP2C125 | GLYMA.20G244300 | 5 | 388 | 42 720.79 | 6.87 | 44.45 | -0.222 | ||||
| GmPP2C126 | GLYMA.20G248900 | 3 | 365 | 39 896.10 | 7.16 | 37.52 | -0.239 | ||||
表2 大豆PP2C家族成员的理化性质
Table 2 Soybean PP2C family members identified in this study
| 基因名 Gene name | 基因登录号 Gene ID | 外显子数量 Exon count | 氨基酸残基 Peptide residues | 分子量 Molecular weight/ku | 等电点 pI | 不稳定系数 Instability index | 亲水性 Grand average of hydropathicity | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| GmPP2C1 | GLYMA.01G127600 | 4 | 290 | 32 192.77 | 6.65 | 37.11 | -0.267 | ||||
| GmPP2C2 | GLYMA.01G158000 | 4 | 374 | 40 365.95 | 5.42 | 55.69 | -0.200 | ||||
| GmPP2C3 | GLYMA.01G191300 | 4 | 377 | 41 796.69 | 6.43 | 40.61 | -0.200 | ||||
| GmPP2C4 | GLYMA.01G199600 | 4 | 753 | 83 208.77 | 5.21 | 50.37 | -0.575 | ||||
| GmPP2C5 | GLYMA.01G225100 | 3 | 324 | 35 093.05 | 6.75 | 47.55 | -0.228 | ||||
| GmPP2C6 | GLYMA.02G009100 | 4 | 396 | 43 461.91 | 4.78 | 51.36 | -0.213 | ||||
| GmPP2C7 | GLYMA.02G044700 | 5 | 394 | 43 795.09 | 8.79 | 47.28 | -0.250 | ||||
| GmPP2C8 | GLYMA.02G058600 | 4 | 364 | 39 141.87 | 5.69 | 34.82 | -0.152 | ||||
| GmPP2C9 | GLYMA.02G144500 | 22 | 1 073 | 121 060.70 | 5.77 | 38.93 | -0.383 | ||||
| GmPP2C10 | GLYMA.02G228200 | 4 | 389 | 41 812.92 | 6.28 | 55.94 | -0.294 | ||||
| GmPP2C11 | GLYMA.02G250200 | 4 | 407 | 44 933.13 | 6.67 | 53.82 | -0.361 | ||||
| GmPP2C12 | GLYMA.03G043100 | 5 | 211 | 23 690.01 | 7.67 | 37.86 | -0.332 | ||||
| GmPP2C13 | GLYMA.03G044100 | 3 | 222 | 24 615.08 | 6.43 | 40.96 | -0.285 | ||||
| GmPP2C14 | GLYMA.03G176100 | 5 | 357 | 39 901.23 | 6.84 | 41.18 | -0.287 | ||||
| GmPP2C15 | GLYMA.03G225800 | 4 | 887 | 99 448.89 | 6.01 | 49.22 | -0.544 | ||||
| GmPP2C16 | GLYMA.03G233300 | 8 | 426 | 45 561.23 | 7.98 | 38.38 | -0.074 | ||||
| GmPP2C17 | GLYMA.03G233500 | 8 | 371 | 41 097.80 | 6.64 | 39.60 | -0.246 | ||||
| GmPP2C18 | GLYMA.04G038000 | 11 | 429 | 46 556.83 | 5.28 | 41.90 | -0.288 | ||||
| GmPP2C19 | GLYMA.04G053800 | 6 | 544 | 59 216.90 | 4.86 | 43.36 | -0.216 | ||||
| GmPP2C20 | GLYMA.04G059000 | 9 | 312 | 33 540.32 | 5.02 | 37.01 | -0.258 | ||||
| GmPP2C21 | GLYMA.04G069900 | 4 | 370 | 40 592.64 | 5.29 | 58.10 | -0.389 | ||||
| GmPP2C22 | GLYMA.04G101400 | 6 | 301 | 33 228.71 | 7.70 | 41.29 | -0.313 | ||||
| GmPP2C23 | GLYMA.05G113400 | 4 | 696 | 76 424.07 | 5.57 | 45.86 | -0.371 | ||||
| GmPP2C24 | GLYMA.05G116800 | 6 | 282 | 30 673.49 | 6.66 | 38.30 | -0.357 | ||||
| GmPP2C25 | GLYMA.05G227100 | 4 | 384 | 42 155.30 | 5.36 | 67.38 | -0.348 | ||||
| GmPP2C26 | GLYMA.06G016100 | 4 | 385 | 41 908.80 | 5.28 | 52.86 | -0.179 | ||||
| GmPP2C27 | GLYMA.06G039000 | 9 | 429 | 46 694.27 | 5.29 | 41.02 | -0.230 | ||||
| GmPP2C28 | GLYMA.06G050900 | 5 | 343 | 37 342.22 | 5.35 | 33.80 | -0.234 | ||||
| GmPP2C29 | GLYMA.06G054000 | 6 | 544 | 58 934.53 | 4.83 | 45.69 | -0.182 | ||||
| GmPP2C30 | GLYMA.06G059700 | 9 | 314 | 33 766.48 | 5.18 | 40.20 | -0.327 | ||||
| GmPP2C31 | GLYMA.06G071900 | 5 | 370 | 40 595.79 | 5.30 | 56.73 | -0.365 | ||||
| GmPP2C32 | GLYMA.06G103100 | 6 | 293 | 32 415.53 | 7.09 | 41.39 | -0.432 | ||||
| GmPP2C33 | GLYMA.06G130900 | 11 | 392 | 43 150.91 | 5.84 | 50.68 | -0.155 | ||||
| GmPP2C34 | GLYMA.06G238200 | 5 | 282 | 30 849.97 | 5.87 | 41.78 | -0.313 | ||||
| GmPP2C35 | GLYMA.06G296200 | 5 | 471 | 51 859.97 | 5.50 | 43.70 | -0.504 | ||||
| GmPP2C36 | GLYMA.07G131800 | 4 | 602 | 66 123.01 | 5.13 | 38.86 | -0.359 | ||||
| GmPP2C37 | GLYMA.07G233000 | 2 | 386 | 42 549.56 | 5.04 | 61.31 | -0.056 | ||||
| GmPP2C38 | GLYMA.07G239000 | 5 | 374 | 41 517.38 | 7.81 | 35.89 | -0.192 | ||||
| GmPP2C39 | GLYMA.07G244700 | 7 | 367 | 41 031.05 | 8.29 | 30.04 | -0.273 | ||||
| GmPP2C40 | GLYMA.07G248000 | 5 | 520 | 56 834.23 | 4.99 | 41.85 | -0.192 | ||||
| GmPP2C41 | GLYMA.08G033800 | 4 | 385 | 42 285.55 | 5.26 | 63.59 | -0.343 | ||||
| GmPP2C42 | GLYMA.08G071900 | 7 | 282 | 30 694.42 | 6.17 | 42.66 | -0.362 | ||||
| GmPP2C43 | GLYMA.08G081400 | 8 | 351 | 39 455.77 | 4.86 | 43.14 | -0.695 | ||||
| GmPP2C44 | GLYMA.08G178800 | 5 | 280 | 30 455.45 | 6.25 | 34.28 | -0.323 | ||||
| GmPP2C45 | GLYMA.09G031700 | 2 | 405 | 44 584.63 | 5.01 | 61.41 | -0.104 | ||||
| GmPP2C46 | GLYMA.09G034800 | 4 | 374 | 41 362.15 | 7.25 | 44.03 | -0.129 | ||||
| GmPP2C47 | GLYMA.09G045100 | 6 | 409 | 45 167.96 | 4.84 | 38.69 | -0.224 | ||||
| GmPP2C48 | GLYMA.09G066500 | 5 | 522 | 57 118.04 | 5.42 | 42.20 | -0.545 | ||||
| GmPP2C49 | GLYMA.09G096600 | 4 | 381 | 42 730.21 | 5.04 | 44.72 | -0.384 | ||||
| GmPP2C50 | GLYMA.09G111300 | 5 | 385 | 43 080.32 | 8.71 | 46.63 | -0.205 | ||||
| GmPP2C51 | GLYMA.09G249700 | 5 | 489 | 54 209.69 | 5.53 | 46.34 | -0.556 | ||||
| GmPP2C52 | GLYMA.09G280700 | 5 | 434 | 48 642.41 | 5.11 | 42.79 | -0.504 | ||||
| GmPP2C53 | GLYMA.10G004200 | 5 | 282 | 32 656.61 | 9.57 | 47.97 | -0.498 | ||||
| GmPP2C54 | GLYMA.10G009800 | 5 | 396 | 43 572.14 | 4.76 | 49.32 | -0.154 | ||||
| GmPP2C55 | GLYMA.10G047500 | 5 | 371 | 40 953.34 | 6.43 | 42.05 | -0.316 | ||||
| GmPP2C56 | GLYMA.10G149600 | 8 | 428 | 45 996.65 | 7.49 | 37.66 | -0.149 | ||||
| GmPP2C57 | GLYMA.10G150100 | 7 | 368 | 41 126.43 | 5.78 | 39.61 | -0.360 | ||||
| GmPP2C58 | GLYMA.10G182400 | 6 | 277 | 31 868.60 | 9.17 | 54.09 | -0.455 | ||||
| GmPP2C59 | GLYMA.10G259400 | 4 | 378 | 42 452.47 | 8.13 | 42.60 | -0.233 | ||||
| GmPP2C60 | GLYMA.10G271500 | 11 | 431 | 46 614.22 | 7.99 | 38.89 | -0.240 | ||||
| GmPP2C61 | GLYMA.10G282900 | 5 | 397 | 43 516.33 | 7.28 | 51.12 | -0.222 | ||||
| GmPP2C62 | GLYMA.10G291800 | 9 | 320 | 34 490.27 | 5.02 | 41.37 | -0.275 | ||||
| GmPP2C63 | GLYMA.10G294400 | 5 | 389 | 43 106.25 | 8.10 | 46.29 | -0.246 | ||||
| GmPP2C64 | GLYMA.10G297900 | 2 | 208 | 22 659.85 | 5.57 | 32.40 | 0.014 | ||||
| GmPP2C65 | GLYMA.11G018000 | 4 | 336 | 36 484.61 | 7.89 | 45.57 | -0.240 | ||||
| GmPP2C66 | GLYMA.11G042300 | 4 | 739 | 82 047.64 | 5.28 | 53.20 | -0.576 | ||||
| GmPP2C67 | GLYMA.11G050900 | 4 | 375 | 41 826.79 | 5.83 | 40.09 | -0.194 | ||||
| GmPP2C68 | GLYMA.11G087000 | 4 | 374 | 40 313.84 | 5.51 | 56.60 | -0.100 | ||||
| GmPP2C69 | GLYMA.11G139500 | 4 | 720 | 79 617.51 | 5.53 | 36.87 | -0.449 | ||||
| GmPP2C70 | GLYMA.11G170200 | 4 | 335 | 36 549.43 | 6.71 | 51.93 | -0.293 | ||||
| GmPP2C71 | GLYMA.11G171600 | 4 | 336 | 36 663.53 | 6.71 | 51.81 | -0.302 | ||||
| GmPP2C72 | GLYMA.11G222600 | 4 | 401 | 43 834.33 | 5.51 | 56.39 | -0.438 | ||||
| GmPP2C73 | GLYMA.12G063100 | 4 | 720 | 79 940.84 | 5.37 | 40.75 | -0.482 | ||||
| GmPP2C74 | GLYMA.12G109500 | 5 | 471 | 51 711.72 | 5.30 | 42.12 | -0.494 | ||||
| GmPP2C75 | GLYMA.12G116800 | 5 | 281 | 31 172.50 | 6.07 | 39.18 | -0.359 | ||||
| GmPP2C76 | GLYMA.12G202400 | 7 | 474 | 51 766.93 | 5.04 | 45.62 | -0.392 | ||||
| GmPP2C77 | GLYMA.12G226100 | 5 | 265 | 29 350.15 | 6.51 | 53.65 | -0.459 | ||||
| GmPP2C78 | GLYMA.13G042100 | 11 | 356 | 39 260.01 | 5.17 | 42.38 | -0.346 | ||||
| GmPP2C79 | GLYMA.13G106800 | 6 | 536 | 58 563.88 | 5.07 | 47.95 | -0.386 | ||||
| GmPP2C80 | GLYMA.13G135300 | 5 | 371 | 40 947.29 | 6.47 | 40.61 | -0.342 | ||||
| GmPP2C81 | GLYMA.13G165300 | 4 | 383 | 42 454.17 | 5.18 | 49.88 | -0.364 | ||||
| GmPP2C82 | GLYMA.13G274800 | 5 | 283 | 31 107.25 | 7.70 | 47.17 | -0.433 | ||||
| GmPP2C83 | GLYMA.13G299300 | 5 | 475 | 52 153.36 | 5.31 | 46.32 | -0.446 | ||||
| GmPP2C84 | GLYMA.14G066400 | 4 | 400 | 44 150.19 | 6.05 | 60.37 | -0.345 | ||||
| GmPP2C85 | GLYMA.14G082200 | 11 | 428 | 46 492.90 | 5.29 | 42.01 | -0.276 | ||||
| GmPP2C86 | GLYMA.14G104900 | 9 | 338 | 36 587.85 | 5.15 | 44.51 | -0.193 | ||||
| GmPP2C87 | GLYMA.14G108400 | 5 | 557 | 60 815.60 | 4.62 | 41.13 | -0.205 | ||||
| GmPP2C88 | GLYMA.14G161400 | 10 | 356 | 39 293.99 | 5.47 | 41.18 | -0.426 | ||||
| GmPP2C89 | GLYMA.14G162100 | 3 | 386 | 42 074.51 | 6.73 | 46.77 | -0.333 | ||||
| GmPP2C90 | GLYMA.14G195200 | 4 | 390 | 41 811.95 | 6.11 | 55.12 | -0.261 | ||||
| GmPP2C91 | GLYMA.15G053700 | 5 | 278 | 30 487.53 | 6.31 | 35.14 | -0.282 | ||||
| GmPP2C92 | GLYMA.15G139500 | 4 | 372 | 41 084.92 | 7.69 | 46.14 | -0.123 | ||||
| GmPP2C93 | GLYMA.15G172500 | 8 | 527 | 57 849.70 | 5.22 | 44.75 | -0.541 | ||||
| GmPP2C94 | GLYMA.15G201300 | 4 | 379 | 42 328.00 | 5.46 | 45.59 | -0.353 | ||||
| GmPP2C95 | GLYMA.16G122200 | 6 | 394 | 43 876.13 | 8.79 | 47.20 | -0.263 | ||||
| GmPP2C96 | GLYMA.17G026100 | 5 | 524 | 57 138.86 | 5.52 | 37.84 | -0.167 | ||||
| GmPP2C97 | GLYMA.17G029200 | 7 | 376 | 41 908.18 | 7.99 | 29.58 | -0.233 | ||||
| GmPP2C98 | GLYMA.17G034500 | 4 | 375 | 41 437.23 | 7.80 | 33.16 | -0.208 | ||||
| GmPP2C99 | GLYMA.17G038000 | 2 | 380 | 41 708.68 | 5.09 | 63.88 | -0.022 | ||||
| GmPP2C100 | GLYMA.17G052500 | 7 | 538 | 58 548.81 | 4.97 | 48.37 | -0.374 | ||||
| GmPP2C101 | GLYMA.17G105800 | 4 | 383 | 42 290.77 | 5.11 | 47.42 | -0.344 | ||||
| GmPP2C102 | GLYMA.17G153800 | 5 | 701 | 77 629.50 | 5.50 | 49.90 | -0.447 | ||||
| GmPP2C103 | GLYMA.17G218700 | 8 | 558 | 60 792.54 | 4.62 | 38.16 | -0.208 | ||||
| GmPP2C104 | GLYMA.17G221100 | 9 | 338 | 36 655.97 | 5.09 | 41.29 | -0.188 | ||||
| GmPP2C105 | GLYMA.17G230600 | 5 | 344 | 37 627.48 | 5.07 | 31.99 | -0.240 | ||||
| GmPP2C106 | GLYMA.17G243100 | 12 | 428 | 46 575.99 | 5.23 | 45.89 | -0.294 | ||||
| GmPP2C107 | GLYMA.18G035000 | 4 | 400 | 44 067.66 | 5.17 | 58.50 | -0.443 | ||||
| GmPP2C108 | GLYMA.18G061000 | 4 | 347 | 37 640.64 | 5.81 | 51.88 | -0.222 | ||||
| GmPP2C109 | GLYMA.18G180400 | 4 | 597 | 65 974.13 | 5.37 | 37.54 | -0.349 | ||||
| GmPP2C110 | GLYMA.18G208100 | 5 | 424 | 47 458.29 | 5.24 | 45.39 | -0.473 | ||||
| GmPP2C111 | GLYMA.18G242700 | 6 | 487 | 54 026.40 | 6.10 | 42.66 | -0.578 | ||||
| GmPP2C112 | GLYMA.18G283300 | 6 | 455 | 50 115.64 | 5.30 | 37.19 | -0.325 | ||||
| GmPP2C113 | GLYMA.19G069200 | 3 | 377 | 40 769.73 | 5.68 | 51.67 | -0.354 | ||||
| GmPP2C114 | GLYMA.19G147800 | 4 | 391 | 44 150.56 | 7.69 | 53.67 | -0.278 | ||||
| GmPP2C115 | GLYMA.19G176600 | 5 | 370 | 41 129.64 | 8.22 | 40.69 | -0.272 | ||||
| GmPP2C116 | GLYMA.19G222700 | 4 | 887 | 99 137.72 | 6.01 | 49.65 | -0.516 | ||||
| GmPP2C117 | GLYMA.19G230200 | 8 | 429 | 46 091.00 | 7.97 | 36.52 | -0.092 | ||||
| GmPP2C118 | GLYMA.19G230600 | 9 | 369 | 40 812.38 | 6.09 | 35.91 | -0.243 | ||||
| GmPP2C119 | GLYMA.20G106800 | 6 | 397 | 43 657.42 | 7.28 | 50.98 | -0.270 | ||||
| GmPP2C120 | GLYMA.20G119100 | 12 | 431 | 46 513.07 | 7.56 | 38.39 | -0.235 | ||||
| GmPP2C121 | GLYMA.20G131500 | 4 | 373 | 41 357.11 | 6.93 | 40.87 | -0.180 | ||||
| GmPP2C122 | GLYMA.20G207600 | 6 | 277 | 31 730.49 | 9.44 | 51.96 | -0.467 | ||||
| GmPP2C123 | GLYMA.20G238100 | 8 | 367 | 41 147.49 | 6.14 | 41.32 | -0.389 | ||||
| GmPP2C124 | GLYMA.20G238700 | 8 | 428 | 46 223.91 | 6.91 | 37.92 | -0.181 | ||||
| GmPP2C125 | GLYMA.20G244300 | 5 | 388 | 42 720.79 | 6.87 | 44.45 | -0.222 | ||||
| GmPP2C126 | GLYMA.20G248900 | 3 | 365 | 39 896.10 | 7.16 | 37.52 | -0.239 | ||||
图2 大豆与拟南芥PP2C家族系统进化分析 进化树中包含126个大豆PP2C家族成员,80个拟南芥PP2C家族成员,共分为11个亚家族。
Fig.2 Phylogenetic tree of PP2C proteins from soybean and Arabidopsis Evolutionary tree contained 126 soybean PP2C family members and 80 Arabidopsis PP2C family members, which were divided into 11 subfamilies.
图4 大豆响应盐胁迫的转录组分析与PP2C家族基因的表达模式 a,对照和盐处理样品的相关分析;b,差异基因数量统计;c,大豆PP2C基因在盐处理前后的表达模式;d,9个差异基因的qRT-PCR结果。
Fig.4 Transcriptome analysis of soybean response to salt stress and expression patterns of PP2C family genes a, Correlation analysis of control and salt-treated samples; b,Statistics of the number of differentially expressed genes; c,Expression pattern of soybean PP2C gene before and after salt treatment; d, qRT-PCR verification of nine differentially expressed genes.
图5 大豆PP2C家族中差异基因的GO富集分析 MF,分子功能;CC,细胞组分;右边的数值为Q值。
Fig.5 GO enrichment analysis of differentially expressed genes in PP2C family of soybeans MF, Molecular function; CC, Cellular component;The value on the right is the Q value.
图6 PP2C蛋白家族互作预测与酵母双杂交分析 a,蛋白互作预测网络图;b、c,酵母双杂交验证蛋白互作,SD/-L/-T为缺失亮氨酸和色氨酸的酵母培养基,SD/-L/-T/-H为缺失亮氨酸、色氨酸和组氨酸的酵母培养基。
Fig.6 Prediction of PP2C protein family interaction and yeast two-hybrid analysis a,Protein interaction prediction network diagram; b and c, yeast two-hybrid verification protein interaction, SD/-L/-T represented yeast culture medium lacking amino acids leucine and tryptophan, SD/-L/-T/-H representedyeast culture medium lacking amino acids leucine, tryptophan and histidine.
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