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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (9): 2179-2186.doi: 10.3724/SP.J.1006.2024.44030

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Identification, evaluation and screening of salt-tolerant of soybean germplasm resources at seedling stage

SUN Xian-Jun1(), HU Zheng1, JIANG Xue-Min2, WANG Shi-Jia2, CHEN Xiang-Qian2, ZHANG Hui-Yuan1, ZHANG Hui1,*(), JIANG Qi-Yan2,*()   

  1. 1Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / State Key Laboratory of Crop Gene Resources and Breeding, Beijing 100081, China
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / Key Laboratory of Grain Crop Genetic Resources Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Beijing 100081, China
  • Received:2024-02-21 Accepted:2024-05-21 Online:2024-09-12 Published:2024-06-05
  • Contact: *E-mail: jiangqiyan@caas.cn; E-mail: zhanghui06@caas.cn
  • Supported by:
    National Key Research and Development Program of China(2021YFD1201603);Key Research and Development Program of Shandong Province(ZDYF2023LZGC001);Key Research and Development Program of Hainan Province(ZDYF2022XDNY135);Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(ASTIP CAAS ZDRW202201);Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(01-ICS-02);National Natural Science Foundation of China(31601329);Collaborative Innovation Center for Modern Crop Production co-sponsored by Province and Ministry(CIC-MCP)

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Abstract:

Soil salinization has a significant impact on soybean quality and yield. Therefore, it is crucial to screen salt-tolerant soybean resources for the comprehensive utilization of salinized land. To establish a method for identifying and evaluating salt tolerance in soybean at the seedling stage, we conducted experiments using fresh water and 10 equal-differential gradients of NaCl solutions ranging from 0.9% to 1.8%. These solutions were applied to vermiculite culture medium when the soybean plants reached the stage of two emerged leaves. Results indicated that the 1.2% salinity treatment for 16 days demonstrated the highest interquartile range of salt-tolerant grading among various soybean germplasm resources, thus considered the optimal condition for evaluating salt tolerance in soybean seedlings. A total of 504 soybean germplasm resources from diverse geographic regions at home and abroad were subjected to evaluation using the soybean salt-tolerant evaluation system. Among them, 46, 146, 157, 79, and 76 soybean germplasm resources received salt-tolerant gradings of 1, 2, 3, 4, and 5, respectively. The soybean resources with salt-tolerant grading 1 were further analyzed using the GmSALT3 gene molecular marker. The amplification results of 40 soybean resources (80.96%) were consistent with the GmSALT3 gene molecular marker. To analyze the change in salt stress concentration in the culture medium during the identification of soybean seedlings, a regression equation was established between soil salt content (Y, %) and electrical conductivity (X, mS cm-1): Y=0.278X-0.0618, with a prediction accuracy above 95%. The variation trend of salt content in the culture medium was measured from the beginning of salt treatment until the end of the investigation, and it was found to be maintained at approximately 13 mS cm-1. This study not only provides a technical system for large-scale identification of salt-tolerant soybean seedlings but also serves as a foundation for breeding new soybean salt-tolerant germplasm resources.

Key words: soybean germplasm resources, salt-tolerant system at seedling stage, salt-tolerant germplasm resources at seedling stage, soil salinity determination

Table S1

The materials and salt-tolerent grade of soybean for evaluation of salt-tolerant at seedling stage"

编号
No.		 统一编号
Unified No.		 耐盐等级
Salt-tolerant grading		 编号
No.		 统一编号
Unified No.		 耐盐等级
Salt-tolerant grading		 编号
No.		 统一编号
Unified No.		 耐盐等级
Salt-tolerant grading
S001 WDD00406 1 S169 ZDD24035 2 S337 ZDD24865 3
S002 WDD00426 1 S170 ZDD24052 2 S338 ZDD24992 3
S003 WDD00731 1 S171 ZDD24072 2 S339 ZDD24998 3
S004 WDD00743 1 S172 ZDD24073 2 S340 ZDD25036 3
S005 WDD01211 1 S173 ZDD24206 2 S341 ZDD25148 3
S006 WDD01609 1 S174 ZDD24208 2 S342 ZDD25158 3
S007 WDD01631 1 S175 ZDD24620 2 S343 ZDD25163 3
S008 WDD01639 1 S176 ZDD24643 2 S344 ZDD25170 3
S009 WDD01646 1 S177 ZDD24742 2 S345 ZDD25181 3
S010 ZDD02145 1 S178 ZDD24761 2 S346 ZDD25192 3
S011 ZDD02587 1 S179 ZDD24795 2 S347 ZDD25193 3
S012 ZDD02858 1 S180 ZDD24840 2 S348 ZDD25214 3
S013 ZDD02911 1 S181 ZDD24971 2 S349 ZDD25353 3
S014 ZDD03036 1 S182 ZDD24980 2 S350 WDD00389 4
S015 ZDD03074 1 S183 ZDD24986 2 S351 WDD00518 4
S016 ZDD03131 1 S184 ZDD24994 2 S352 WDD00534 4
S017 ZDD03412 1 S185 ZDD25018 2 S353 WDD00634 4
S018 ZDD03532 1 S186 ZDD25045 2 S354 WDD00671 4
S019 ZDD04094 1 S187 ZDD25060 2 S355 WDD00799 4
S020 ZDD04302 1 S188 ZDD25080 2 S356 WDD00950 4
S021 ZDD07262 1 S189 ZDD25084 2 S357 WDD00970 4
S022 ZDD07639 1 S190 ZDD25178 2 S358 WDD01043 4
S023 ZDD08002 1 S191 ZDD25222 2 S359 WDD01153 4
S024 ZDD08536 1 S192 ZDD25267 2 S360 WDD01242 4
S025 ZDD09897 1 S193 WDD00457 3 S361 WDD01563 4
S026 ZDD11265 1 S194 WDD00553 3 S362 WDD02023 4
S027 ZDD11434 1 S195 WDD00691 3 S363 WDD02208 4
S028 ZDD11763 1 S196 WDD00791 3 S364 ZDD00070 4
S029 ZDD12732 1 S197 WDD00816 3 S365 ZDD00749 4
S030 ZDD13930 1 S198 WDD00837 3 S366 ZDD01652 4
S031 ZDD18574 1 S199 WDD00973 3 S367 ZDD01670 4
S032 ZDD19787 1 S200 WDD00975 3 S368 ZDD02169 4
S033 ZDD19872 1 S201 WDD01010 3 S369 ZDD02271 4
S034 ZDD19976 1 S202 WDD01117 3 S370 ZDD02547 4
S035 ZDD20809 1 S203 WDD01209 3 S371 ZDD02742 4
S036 ZDD21644 1 S204 WDD01210 3 S372 ZDD03556 4
S037 ZDD23044 1 S205 WDD01219 3 S373 ZDD03712 4
S038 ZDD23113 1 S206 WDD01238 3 S374 ZDD03735 4
S039 ZDD23208 1 S207 WDD01256 3 S375 ZDD04393 4
S040 ZDD23298 1 S208 WDD01365 3 S376 ZDD05534 4
S041 ZDD23324 1 S209 WDD01366 3 S377 ZDD05540 4
S042 ZDD23325 1 S210 WDD01469 3 S378 ZDD05960 4
S043 ZDD23335 1 S211 WDD01634 3 S379 ZDD08181 4
S044 ZDD23528 1 S212 WDD01885 3 S380 ZDD09494 4
S045 ZDD25046 1 S213 WDD02017 3 S381 ZDD10081 4
S046 ZDD25171 1 S214 WDD02350 3 S382 ZDD10481 4
S047 WDD00393 2 S215 WDD03088 3 S383 ZDD10566 4
S048 WDD00404 2 S216 ZDD00137 3 S384 ZDD10628 4
S049 WDD00448 2 S217 ZDD00239 3 S385 ZDD10836 4
S050 WDD00636 2 S218 ZDD00404 3 S386 ZDD10870 4
S051 WDD00734 2 S219 ZDD00412 3 S387 ZDD11089 4
S052 WDD00750 2 S220 ZDD00583 3 S388 ZDD11458 4
S053 WDD00761 2 S221 ZDD00948 3 S389 ZDD11635 4
S054 WDD00926 2 S222 ZDD01645 3 S390 ZDD11656 4
S055 WDD00992 2 S223 ZDD02253 3 S391 ZDD12076 4
S056 WDD00994 2 S224 ZDD02304 3 S392 ZDD12427 4
S057 WDD01006 2 S225 ZDD02325 3 S393 ZDD12893 4
S058 WDD01007 2 S226 ZDD02326 3 S394 ZDD12894 4
S059 WDD01200 2 S227 ZDD02327 3 S395 ZDD13160 4
S060 WDD01206 2 S228 ZDD02342 3 S396 ZDD13172 4
S061 WDD01273 2 S229 ZDD02356 3 S397 ZDD13339 4
S062 WDD01576 2 S230 ZDD02368 3 S398 ZDD14154 4
S063 WDD01649 2 S231 ZDD02452 3 S399 ZDD14206 4
S064 WDD01872 2 S232 ZDD02471 3 S400 ZDD14533 4
S065 WDD01903 2 S233 ZDD02475 3 S401 ZDD14534 4
S066 WDD01959 2 S234 ZDD02479 3 S402 ZDD15516 4
S067 WDD02125 2 S235 ZDD02483 3 S403 ZDD16095 4
S068 WDD02138 2 S236 ZDD02486 3 S404 ZDD16331 4
S069 WDD02283 2 S237 ZDD02496 3 S405 ZDD16358 4
S070 WDD03111 2 S238 ZDD02565 3 S406 ZDD16810 4
S071 ZDD00215 2 S239 ZDD02580 3 S407 ZDD16842 4
S072 ZDD00444 2 S240 ZDD02791 3 S408 ZDD17188 4
S073 ZDD01535 2 S241 ZDD03786 3 S409 ZDD17629 4
S074 ZDD02151 2 S242 ZDD04296 3 S410 ZDD19457 4
S075 ZDD02197 2 S243 ZDD05179 3 S411 ZDD19992 4
S076 ZDD02501 2 S244 ZDD05388 3 S412 ZDD20086 4
S077 ZDD02963 2 S245 ZDD05426 3 S413 ZDD20100 4
S078 ZDD03102 2 S246 ZDD05473 3 S414 ZDD20157 4
S079 ZDD03375 2 S247 ZDD05503 3 S415 ZDD20214 4
S080 ZDD03417 2 S248 ZDD05570 3 S416 ZDD20531 4
S081 ZDD03530 2 S249 ZDD05622 3 S417 ZDD20912 4
S082 ZDD03598 2 S250 ZDD05631 3 S418 ZDD20954 4
S083 ZDD03653 2 S251 ZDD06258 3 S419 ZDD20972 4
S084 ZDD03660 2 S252 ZDD06490 3 S420 ZDD21399 4
S085 ZDD03686 2 S253 ZDD08000 3 S421 ZDD21927 4
S086 ZDD03808 2 S254 ZDD08172 3 S422 ZDD23316 4
S087 ZDD03831 2 S255 ZDD08359 3 S423 ZDD23399 4
S088 ZDD03835 2 S256 ZDD08553 3 S424 ZDD24034 4
S089 ZDD03836 2 S257 ZDD08656 3 S425 ZDD24542 4
S090 ZDD03895 2 S258 ZDD08890 3 S426 ZDD25075 4
S091 ZDD04063 2 S259 ZDD08950 3 S427 ZDD25183 4
S092 ZDD04163 2 S260 ZDD08988 3 S428 ZDD25343 4
S093 ZDD04301 2 S261 ZDD09106 3 S429 WDD00373 5
S094 ZDD04362 2 S262 ZDD09121 3 S430 WDD01021 5
S095 ZDD04417 2 S263 ZDD09203 3 S431 ZDD00110 5
S096 ZDD04681 2 S264 ZDD09254 3 S432 ZDD00247 5
S097 ZDD05444 2 S265 ZDD09259 3 S433 ZDD00328 5
S098 ZDD05585 2 S266 ZDD09372 3 S434 ZDD00359 5
S099 ZDD06112 2 S267 ZDD09398 3 S435 ZDD00360 5
S100 ZDD06996 2 S268 ZDD09451 3 S436 ZDD00373 5
S101 ZDD07660 2 S269 ZDD10503 3 S437 ZDD00565 5
S102 ZDD07905 2 S270 ZDD10766 3 S438 ZDD00818 5
S103 ZDD07927 2 S271 ZDD10823 3 S439 ZDD00971 5
S104 ZDD08040 2 S272 ZDD10914 3 S440 ZDD01472 5
S105 ZDD08102 2 S273 ZDD10921 3 S441 ZDD01620 5
S106 ZDD08199 2 S274 ZDD10970 3 S442 ZDD01626 5
S107 ZDD08532 2 S275 ZDD11267 3 S443 ZDD04029 5
S108 ZDD09856 2 S276 ZDD11279 3 S444 ZDD04855 5
S109 ZDD09979 2 S277 ZDD11336 3 S445 ZDD05634 5
S110 ZDD10160 2 S278 ZDD11450 3 S446 ZDD05923 5
S111 ZDD10549 2 S279 ZDD11543 3 S447 ZDD05928 5
S112 ZDD10563 2 S280 ZDD11596 3 S448 ZDD05930 5
S113 ZDD10581 2 S281 ZDD11614 3 S449 ZDD05935 5
S114 ZDD10637 2 S282 ZDD12366 3 S450 ZDD06924 5
S115 ZDD10920 2 S283 ZDD12623 3 S451 ZDD07448 5
S116 ZDD10994 2 S284 ZDD12866 3 S452 ZDD08001 5
S117 ZDD11233 2 S285 ZDD13283 3 S453 ZDD08004 5
S118 ZDD11268 2 S286 ZDD14162 3 S454 ZDD08047 5
S119 ZDD11281 2 S287 ZDD14167 3 S455 ZDD08091 5
S120 ZDD11352 2 S288 ZDD14174 3 S456 ZDD08189 5
S121 ZDD11354 2 S289 ZDD14518 3 S457 ZDD08197 5
S122 ZDD11509 2 S290 ZDD14521 3 S458 ZDD08244 5
S123 ZDD11577 2 S291 ZDD14700 3 S459 ZDD10456 5
S124 ZDD11582 2 S292 ZDD14711 3 S460 ZDD10589 5
S125 ZDD11620 2 S293 ZDD16055 3 S461 ZDD10595 5
S126 ZDD11925 2 S294 ZDD17649 3 S462 ZDD10888 5
S127 ZDD12071 2 S295 ZDD18341 3 S463 ZDD11690 5
S128 ZDD12889 2 S296 ZDD18846 3 S464 ZDD11737 5
S129 ZDD14525 2 S297 ZDD18882 3 S465 ZDD12271 5
S130 ZDD14705 2 S298 ZDD18894 3 S466 ZDD12579 5
S131 ZDD14788 2 S299 ZDD19157 3 S467 ZDD12741 5
S132 ZDD17563 2 S300 ZDD19497 3 S468 ZDD12821 5
S133 ZDD17933 2 S301 ZDD19508 3 S469 ZDD12877 5
S134 ZDD18096 2 S302 ZDD19585 3 S470 ZDD12964 5
S135 ZDD18576 2 S303 ZDD19639 3 S471 ZDD12985 5
S136 ZDD19014 2 S304 ZDD19811 3 S472 ZDD13582 5
S137 ZDD19104 2 S305 ZDD19890 3 S473 ZDD14159 5
S138 ZDD19439 2 S306 ZDD20087 3 S474 ZDD14172 5
S139 ZDD19449 2 S307 ZDD20172 3 S475 ZDD14467 5
S140 ZDD19558 2 S308 ZDD20181 3 S476 ZDD14654 5
S141 ZDD19907 2 S309 ZDD20426 3 S477 ZDD14686 5
S142 ZDD19919 2 S310 ZDD20437 3 S478 ZDD14912 5
S143 ZDD19954 2 S311 ZDD20693 3 S479 ZDD15011 5
S144 ZDD19979 2 S312 ZDD20817 3 S480 ZDD16013 5
S145 ZDD20401 2 S313 ZDD20992 3 S481 ZDD16354 5
S146 ZDD20435 2 S314 ZDD21002 3 S482 ZDD16811 5
S147 ZDD20436 2 S315 ZDD21049 3 S483 ZDD17719 5
S148 ZDD20910 2 S316 ZDD21435 3 S484 ZDD18082 5
S149 ZDD20914 2 S317 ZDD21491 3 S485 ZDD18167 5
S150 ZDD21060 2 S318 ZDD21998 3 S486 ZDD18390 5
S151 ZDD22099 2 S319 ZDD22265 3 S487 ZDD18445 5
S152 ZDD22103 2 S320 ZDD22268 3 S488 ZDD18530 5
S153 ZDD22221 2 S321 ZDD23306 3 S489 ZDD20816 5
S154 ZDD22884 2 S322 ZDD23315 3 S490 ZDD20818 5
S155 ZDD23039 2 S323 ZDD23327 3 S491 ZDD20845 5
S156 ZDD23061 2 S324 ZDD23398 3 S492 ZDD21066 5
S157 ZDD23326 2 S325 ZDD23413 3 S493 ZDD21484 5
S158 ZDD23331 2 S326 ZDD23423 3 S494 ZDD22864 5
S159 ZDD23382 2 S327 ZDD23894 3 S495 ZDD22866 5
S160 ZDD23430 2 S328 ZDD23966 3 S496 ZDD22913 5
S161 ZDD23571 2 S329 ZDD24001 3 S497 ZDD23026 5
S162 ZDD23782 2 S330 ZDD24075 3 S498 ZDD23073 5
S163 ZDD23795 2 S331 ZDD24161 3 S499 ZDD23102 5
S164 ZDD23870 2 S332 ZDD24659 3 S500 ZDD23798 5
S165 ZDD23902 2 S333 ZDD24663 3 S501 ZDD23875 5
S166 ZDD23957 2 S334 ZDD24709 3 S502 ZDD24043 5
S167 ZDD24021 2 S335 ZDD24750 3 S503 ZDD24873 5
S168 ZDD24033 2 S336 ZDD24776 3 S504 ZDD25281 5

Table 1

Reference standard for salt-tolerant grading of soybean at seedling stage"

耐盐级别
Salt-tolerant grading		 大豆植株表型
Symptom of soybean plants
1 生长正常, 基本无受害症状, 植株枯死率0-20.0%。
Normal growth, no symptoms of injury, and a plant senescence rate of 0-20.0%.
2 生长基本正常, 植株枯死率20.1%-40.0%。
The growth was generally normal, with a plant senescence rate of 20.1%-40.0%.
3 生长接近正常, 植株枯死率40.1%-60.0%。
The growth was close to normal, with a plant senescence rate of 40.1%-60.0%.
4 生长受抑制, 植株枯死率60.1%-80.0%。
Growth inhibition, with a plant senescence rate of 60.1%-80.0%.
5 植物死亡或生长严重受阻, 植株枯死率80.1%-100%。
Dead or severely stunted, with a plant senescence rate of 80.1%-100%.

Table 2

Interquartile difference of salt-tolerant grading among resources under different salt concentrations"

NaCl含量
NaCl
content (%)		 1st-5th day 6th
day		 7th
day		 8th
day		 9th
day		 10th day 11th day 12th day 13th day 14th day 15th day 16th day 17th day 18th day 19th day 20th day
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
0.9 0 0 0 0 0 0 0 0 0 0 0 0 0 0.13 0.13 0.63
1.0 0 0 0 0 0 0 0 0 0 0 0.13 0.50 0.50 0.63 1.13 1.13
1.1 0 0 0 0 0 0 0 0 0.50 0.50 0.50 1.00 1.00 1.50 1.50 1.50
1.2 0 0 0 0 0 0 0.13 0.50 0.50 1.00 1.00 1.13 1.00 1.00 1.00 0.63
1.3 0 0 0 0 0 0 0 0.50 0.50 0.13 0.50 0.50 1.00 0.63 0.50 1.00
1.4 0 0 0 0 0.13 0.50 0.50 1.00 0.63 0.50 0.63 0.50 0.50 0.50 0.63 0.50
1.5 0 0 0 0.50 0.63 1.00 1.00 1.00 1.00 1.00 0.63 0.50 0.50 0.50 0.50 0
1.6 0 0 0.63 1.00 1.00 0.50 0.50 0.50 0.50 0.50 0.13 0 0 0 0 0
1.7 0 0 0.63 0.63 1.00 0.50 0.50 0 0 0 0 0 0 0 0 0
1.8 0 0.13 0.50 0.50 0 0 0 0 0 0 0 0 0 0 0 0

Fig. 1

Identification and evaluation of salt-tolerant soybean at seedling stage A: phenotypes of soybean of different salt-tolerant grades at seedling stage; B: statistics of soybean of different salt-tolerant grades at seedling stage."

Fig. 2

Determination of conductivity of soil and vermiculite A: conductivity of supernatant and salt content of soil; B: measurement of conductivity of culture substrate."

Table 3

Analysis of evaluation accuracy of regression equation"

土壤含盐量
Salt content of soil (%)		 电导率
Electric conductivity (mS cm-1)		 预测值
Predicted value (%)		 差值
Difference		 预测精度
Evaluation accuracy (%)
0.0 0.30 0.02 0.02 97.87
0.1 0.69 0.13 0.03 97.08
0.2 0.89 0.19 -0.01 98.59
0.3 1.42 0.33 0.03 96.76
0.4 1.79 0.44 0.04 96.45
0.5 2.13 0.53 0.03 97.08
0.6 2.42 0.61 0.01 99.02
0.7 2.77 0.71 0.01 99.31
0.8 3.23 0.84 0.04 96.33
0.9 3.49 0.91 0.01 99.24
1.0 3.84 1.01 0.01 99.43
1.1 4.20 1.11 0.01 99.42
1.2 4.52 1.20 0.00 99.56
1.3 4.86 1.29 -0.01 99.04
1.4 5.24 1.39 -0.01 99.49
1.5 5.45 1.45 -0.05 95.30

Table S2

The conductivity of culture medium under different treatments (mS cm-1)"

取样时间
Sampling time (d)		 处理1
Treatment 1		 处理2
Treatment 2		 处理3
Treatment 3
1 7.33 8.29 0.51
2 6.42 8.62 0.49
3 7.10 10.73 0.51
4 7.56 12.49 0.52
5 6.22 12.12 0.39
6 5.73 12.15 0.43
7 5.94 13.45 0.56
8 7.12 14.03 0.36
9 8.03 14.46 0.43
10 9.95 13.88 0.35
11 9.55 13.73 0.38
12 10.12 13.70 0.39
13 10.23 12.21 0.39
14 10.33 12.90 0.31
15 11.22 13.21 0.34
16 11.25 12.91 0.22
17 11.08 12.82 0.36
18 11.32 12.07 0.26
19 11.35 12.40 0.28
20 11.29 12.58 0.34
21 11.30 12.25 0.25
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