Welcome to Acta Agronomica Sinica，

Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (3): 427-437.doi: 10.3724/SP.J.1006.2021.04178

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

Efficient screening system of effective sgRNA for cotton CRISPR/Cas9 gene editing

ZHOU Guan-Tong(), LEI Jian-Feng, DAI Pei-Hong, LIU Chao, LI Yue, LIU Xiao-Dong*()   

  1. College of Agriculture, Xinjiang Agricultural University, Engineering Research Centre of Cotton of Ministry of Education, Urumqi 830052, Xinjiang, China
  • Received:2020-08-05 Accepted:2020-10-14 Online:2021-03-12 Published:2020-10-28
  • Contact: LIU Xiao-Dong E-mail:601930485@qq.com;xiaodongliu75@aliyun.com
  • Supported by:
    National Natural Science Foundation of China(31660433);Xinjiang Agricultural University Postgraduate Research and Innovation Project(XJAUGRI2017003)

RichHTML380

29

PDF

1072

Abstract:

Single guide RNA (sgRNA) is one of the important elements of the CRISPR/Cas9 genome editing technology system. However, studies have shown that many sgRNAs cannot work effectively. It is worth screening to verify the effectiveness of multiple design candidate sgRNAs. Instantaneous transformation of protoplasts or leaves with complete editing vectors were used to verification of the effectiveness of sgRNA in the early stage. These methods are time-consuming and laborious, and the success rate is not high, especially for cotton with low efficiency of the protoplasmic system. In this study, target sequences were designed for GhMAPKKK2 and GhAE genes, and two vectors of GhU6-5P::MAPKKK2-sgRNA-1300, GhU6-5P::AE-sgRNA-1300 which transcibed only sgRNA were constructed and injected YZ-1 Cas9 transgenic cotton plant leaves through Agrobacterium; meanwhile, two corresponding complete CRISPR/Cas9 genome editing vectors of GhU6-5P::MAPKKK2-sgRNA-Cas9 and GhU6-5P::AE-sgRNA-Cas9 were constructed and injected YZ-1 wild-type cotton leaves with Agrobacterium. In addition, target sequences were designed for GhPDS, GhCLA1, GhMAPKKK2, and GhAE genes, respectively, and GhU6-5P-2::PDS-sgRNA- CLCrVA, GhU6-5P-2::CLA1-sgRNA-CLCrVA, GhU6-5P-2::MAPKKK2-sgRNA-CLCrVA and GhU6-5P-2::AE-sgRNA-CLCrVA virus delivery vectors were constructed and injected YZ-1 Cas9 transgenic cotton plant leaves through Agrobacterium. In the above experiments, the plants transformed with the empty vector were used as controls. The genomic DNA of the transformed cotton leaves was subjected to PCR and enzyme digestion, and the PCR products which were not completely digested were cloned and sequenced. The results showed that no mutation in target gene was detected in the cotton plants transformed with the GhU6-5P::AE-sgRNA-1300, GhU6-5P::MAPKKK2-sgRNA-Cas9 and GhU6-5P::AE-sgRNA-Cas9, and the target genes mutation in the Cas9 transgenic plants transformed with GhU6-5P::MAPKKK2-sgRNA-1300, GhU6-5P-2::PDS-sgRNA-CLCrVA, GhU6-5P-2::CLA1-sgRNA-CLCrVA, GhU6-5P-2::MAPKKK2-sgRNA-CLCrVA and GhU6-5P-2::AE-sgRNA-CLCrVA vector was uncovered. The types of mutations included base substitution, base deletion and base insertion. The results indicated that the strategy of using Cas9 transgenic plants as transformation recipients can efficiently and truly verify the effectiveness of sgRNA, which eliminated false negative results due to low transformation efficiency, and the strategy of using virus as vectors to deliver sgRNA was more efficient and accurate. The establishment of this sgRNA high-efficiency verification system provides an important technical basis for cotton functional genomics research.

Key words: cotton, transient transformation, CRISPR/Cas9, genome editing

Table 1

Primer sequences used in this study"

引物名称
Primer name		 上游引物序列
Forward sequence (5'-3')		 下游引物序列
Reverse sequence (5'-3')
GhU6-5P-MAPKKK2-sg AAGGGTTCCCAGCTGACATA TATGTCAGCTGGGAACCCTT
GhU6-5P-AE-sg GAGTTTGGAGGGCTTACAAT ATTGTAAGCCCTCCAAACTC
GhU6-5P-2-PDS-sg GAAGCGAGAGATGTTCTAGG CCTAGAACATCTCTCGCTTC
GhU6-5P-2-CLA1-sg TATGCTCGCGGAATGATCAG CTGATCATTCCGCGAGCATA
Test MAPKKK2-sg CCATGTCGTAGCTTATAAAGG TCATTTACCTTCTCTTCCCAG
Test AE-sg AGACTTGTTTCAATGGACTC AATAAGCTGACAGCAGTTGG
Test PDS-sg TGCATGATCCATCACTCAAGTTT GAACGAAAGGCCCTTTCTTTC
Test CLA1-sg GGATCTGAAAGGTGAAAGGAATC TACCGTGATACTTGTCAGCAGCT

Fig. 1

Identification of gene editing vector by restriction enzyme digestion A: 1, 2 are GhU6-5P::MAPKKK2-sgRNA-1300 and GhU6-5P::MAPKKK2-sgRNA-Cas9. B: 1, 2 are GhU6-5P::AE-sgRNA-1300 and GhU6-5P:: AE-sgRNA-Cas9. C: 1-4 are GhU6-5P-2::PDS-sgRNA-ClCrVA , GhU6-5P-2::CLA1-sgRNA-ClCrVA, GhU6-5P-2::MAPKKK2-sgRNA- ClCrVA and GhU6-5P-2::AE-sgRNA-ClCrVA. M: 2K plus II DNA marker."

Fig. 2

GhU6-5P::MAPKKK2-sgRNA targeted mutations A: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion; 3-6: PCR/RE assay of GhU6-5P::MAPKKK2-sgRNA-Cas9 genome of single plant sample after enzyme digestion. B: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and of single plant sample after the enzyme digestion; 3-6: PCR/RE assay of GhU6-5P::MAPKKK2-sgRNA-1300 genome after enzyme digestion. C: sequencing of GhMAPKKK2-sgRNA target sequence and mutant PCR products. M1 occurred at GhMAPKKK2 from D subgenome, M2, M3 occurred at GhMAPKKK2 from A subgenome. D: sequencing peak diagram of the mutant PCR product clone, the red box indicates that the base has a mutation region compared to the control. PCR/RE indicates PCR/restriction enzyme assays."

Fig. 3

GhU6-5P::AE-sgRNA targeted mutations A: M: 2K plus II DNA marker; 12, 11: PCR/RE assay of negative control genome before and after the enzyme digestion; 3-12: PCR/RE assay of GhU6-5P::AE-sgRNA-Cas9 of single plant sample after enzyme digestion. B: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion; 3-12: PCR/RE assay of GhU6-5P::AE-sgRNA-1300 of single plant sample genome after enzyme digestion. PCR/RE indicates PCR/restriction enzyme assays."

Fig. 4

Results of GhU6-5P-2::MAPKKK2-sgRNA-CLCrVA partial editing effect sequencing A: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion. B: M: 2K plus II DNA marker; 3-6: PCR/RE assay of GhU6-5P::MAPKKK2-sgRNA-CLCrVA genome of single plant sample after enzyme digestion. C: M: 2K plus II DNA marker, 1-10: clony PCR product after enzyme digestion. PCR/RE indicates PCR/restriction enzyme assays."

Fig. 5

GhU6-5P-2::AE-sgRNA-CLCrVA partial editing effect sequencing results A: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion. B: M: 2K plus II DNA marker, 3-7: PCR/RE assay of GhU6-5P::AE-sgRNA-CLCrVA genome of single plant sample after enzyme digestion. C: M: 2K plus II DNA marker; 1-10: clony PCR product after enzyme digestion. PCR/RE indicates PCR/restriction enzyme assays."

Fig. 6

Results of GhU6-5P-2::PDS-sgRNA-CLCrVA partial editing effect sequencing A: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome after and before the enzyme digestion. B: M: 2K plus II DNA marker; 3-5: PCR/RE assay of GhU6-5P-2::PDS-sgRNA-CLCrVA of single plant sample genome after enzyme digestion. C: sequencing of GhPDS-sgRNA target sequence and mutant PCR products. M1, M2 occurred at GhPDS from A subgenome; M3 occurred at GhPDS from D subgenome. D: sequencing peak diagram of the mutant PCR product; the red box indicates that the base has a mutation region compared to the control. PCR/RE indicates PCR/restriction enzyme assays."

Fig. 7

Results of GhU6-5P-2::CLA1-sgRNA-CLCrVA partial editing effect sequencing A: M: 2K plus II DNA marker; 1, 2: PCR/RE assay of negative control genome before and after the enzyme digestion. B: M: 2K plus II DNA marker, 3-5: PCR/RE assay of GhU6-5P::CLA1-sgRNA-CLCrVA genome of single plant sample after enzyme digestion. C: sequencing of GhCLA1-sgRNA target sequence and mutant PCR products. M1, M3 occurred at GhCLA1 from D subgenome; M2 occurred at GhCLA1 from A subgenome. D: sequencing peak diagram of the mutant PCR product; the red box indicates that the base has a mutation region compared to the control. PCR/RE indicates PCR/restriction enzyme assays."

[1] 许宗弘. 棉花枯黄萎病研究现状及展望. 知识经济, 2010,16:132.
Xu Z H. Research status and prospect of cotton Fusarium Wilt. Knowledge Econ, 2010,16:132 (in Chinese with English abstract).
[2] 任爱霞. 棉花枯黄萎病抗性遗传及生化机理研究. 浙江大学硕士学位论文, 浙江杭州, 2002.
Ren A X. Study on Inheritance and Biochemical Mechanism of Cotton Fusarium Wilt Resistance. MS Thesis of Zhejiang University, Hangzhou, Zhejiang, China, 2002 (in Chinese with English abstract).
[3] 徐立华. 我国棉花高产、高效栽培技术研究现状与发展思路. 中国棉花, 2001, (3):5-8.
Xu L H. Research status and development ideas of cotton high-yield and high-efficiency cultivation technology in my country. China Cotton, 2001, (3):5-8 (in Chinese with English abstract).
[4] 孙学振, 施培, 周治国. 我国棉花高产栽培技术理论研究现状与展望. 中国棉花, 1999, (4):2-7.
Sun X Z, Shi P, Zhou Z G. Current status and prospects of the theoretical research on cotton high-yield cultivation techniques in my country. China Cotton, 1999, (4):2-7 (in Chinese with English abstract).
[5] Sun Y, Li J, Xia L. Precise genome modification via sequence specific nucleases-mediated gene targetingfor crop improvement. Front Plant Sci, 2016,7:1928.
doi: 10.3389/fpls.2016.01928 pmid: 28066481
[6] 刘蓓, 尉玮, 王丽华. 基因编辑新技术研究进展. 亚热带农业研究, 2013,9(4):262-269.
Liu B, Wei W, Wang L H. Research progress of new technology of gene editing. Subtrop Agric Res, 2013,9(4):262-269 (in Chinese with English abstract).
[7] Cao H X, Wang W, Le H T, Vu G T. The power of CRISPR-Cas9-induced genome editing to speed up plant breeding. Int J Genomics, 2016,2016:5078796.
[8] Gilbert L A, Larson M H, Morsut L, Liu Za, Brar G A, Torres S E, Stern-Ginossar N, Brandman O, Whitehead E H, Doudna J A, Lim W A, Weissman J S, Qi L S. CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Cell, 2013,154:442-451.
[9] Hsu P D, Lander E S, Zhang F. Development and applications of CRISPR-Cas9 for genome engineering. Cell, 2014,157:1262-1278.
pmid: 24906146
[10] Bassett A R, Tibbit C, Ponting C P, Liu J L. Highly efficient targeted mutagenesis of drosophila with the CRISPR/Cas9 system. Cell Rep, 2014,6:1178-1179.
[11] Barrangou R, Marraffini L A. CRISPR-Cas systems: prokaryotes upgrade to adaptive immunity. Mol Cell, 2014,54:234-244.
[12] Mao Y F, Zhang Z J, Feng Z Y, Wei P L, Zhang H, Botella J R, Zhu J K. Development of germline specific CRISPR/Cas9 systems to improve the production of heritable gene modifications in Arabidopsis. Plant Biotechnol J, 2016,14:519-532.
doi: 10.1111/pbi.12468 pmid: 26360626
[13] Kim H, Kim S T, Ryu J, Choi M K, Kweon J, Kang B C, Ahn H M, Bae S, Kim J, Kim J S, Kim S G. A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR/Cas system. J Integr Plant Biol, 2016,58:705-712.
doi: 10.1111/jipb.12474
[14] Gao S L, Tong Y Y, Wen Z Q, Zhu L, Ge M, Chen D J, Jiang Y, Yang S. Multiplex gene editing of theYarrowia lipolytica genome using the CRISPR/Cas9 system. J Ind Microbiol Biotechnol, 2016,43:1085-1093.
pmid: 27349768
[15] Zhang F, Maeder M L, Unger-Wallace E, Hoshaw J P, Reyon D, Christian M, Li X H, Pierick C J, Dobbs D, Peterson T, Joung J K, Voytas D F. High frequency targeted mutagenesis inArabidopsis thaliana using zinc finger nucleases. Proc Natl Acad Sci USA, 2010,107:12028-12033.
doi: 10.1073/pnas.0914991107 pmid: 20508152
[16] Shukla V K, Doyon Y, Miller J C, DeKelver R C, Moehle E A, Worden S E, Mitchell J C, Arnold N L, Gopalan S, Meng X D, Choi V M, Rock J M, Wu Y Y, Katibah G E, Gao Z F, McCaskill D, Simpson M A, Blakeslee B, Greenwalt S A, Butler H J, Hinkley S J, Zhang L, Rebar E J, Gregory P D, Urnov F D. Precise genome modification in the crop speciesZea mays using zinc-finger nucleases. Nature, 2009,459:437-441.
doi: 10.1038/nature07992 pmid: 19404259
[17] Townsend J A, Wright D A, Winfrey R J, Fu F L, Maeder M L, Joung J K, Voytas D F. High-frequency modification of plant genes using engineered zinc-finger nucleases. Nature, 2009,459:442-445.
pmid: 19404258
[18] 谢小东, 高军平, 李泽锋, 张剑锋, 魏攀, 罗朝鹏, 王晨, 武明珠, 翟妞, 杨军. CRISPR/Cas9介导烟草多基因编辑体系的应用. 中国烟草学报, 2019,25(4):72-80.
Xie X D, Gao J P, Li Z F, Zhang J F, Wei P, Luo Z P, Wang C, Wu M Z, Zhai N, Yang J. Application of CRISPR/Cas9 mediated tobacco multi-gene editing system. Acta Tab Sin, 2019,25(4):72-80 (in Chinese with English abstract).
[19] 王海明, 张立强, 李娜, 刘建丰, 马崇烈. 利用CRISPR/Cas9基因编辑技术敲除水稻NRR基因促进根系生长的研究. 杂交水稻, 2019,34(5):39-45.
Wang H M, Zhang L Q, Li N, Liu J F, Ma C L. Using CRISPR/Cas9 gene editing technology to knock out rice NRR gene to promote root growth. Hybrid Rice, 2019,34(5):39-45 (in Chinese with English abstract).
[20] 陈修贵. CRISPR/Cas9系统介导的棉花GhCLA1和GhVP基因编辑的研究. 华中农业大学博士学位论文, 湖北武汉, 2017.
Chen X G. Study on Cotton GhCLA1 and GhVP Gene Editing Mediated by CRISPR/Cas9 System. PhD Dissertation of Huazhong Agricultural University, Wuhan, Hubei, China, 2017 (in Chinese with English abstract).
[21] 王艳玲, 孟志刚, 李妍妍, 孟钊红, 王远, 孙国清, 朱涛, 梁成真, 蔡永萍, 郭三堆, 张锐, 林毅. CRISPR/Cas9编辑棉花精氨酸酶基因促进侧根形成和发育. 中国科学: 生命科学, 2017,47:1200-1203.
Wang Y L, Meng Z G, Li Y Y, Meng Z H, Wang Y, Sun G Q, Zhu T, Liang C Z, Cai Y P, Guo S D, Zhang R, Lin Y. CRISPR/Cas9 editing cotton arginase gene promotes lateral root formation and development. Sci Sin (Vitae), 2017,47:1200-1203 (in Chinese with English abstract).
[22] Farboud B, Meyer B J. Dramatic enhancement of genome editing by CRISPR/Cas9 through improved guide RNA design. Genetics, 2015,199:959-971.
doi: 10.1534/genetics.115.175166 pmid: 25695951
[23] Chen X G, Lu X K, Shu N, Wang S, Wang J J, Wang D L, Guo L X, Ye W W. Targeted mutagenesis in cotton (Gossypium hirsutum L.) using the CRISPR/Cas9 system. Sci Rep, 2017,7:44304.
pmid: 28287154
[24] Gao W, Long L, Tian X Q, Xu F C, Liu J, Prashant K S, Jose R B, Song C P. Genome editing in cotton with the CRISPR/Cas9 system. Front Plant Sci, 2017,8:1364.
[25] Hu J C, Li S, Li Z L, Li H Y, Song W B, Zhao H M, Lai J S, Xia L Q, Li D W, Zhang Y L. A barley stripe mosaic virus-based guide RNA delivery system for targeted mutagenesis in wheat and maize. Mol Plant Pathol, 2019,20:1463-1474.
pmid: 31273916
[26] Yin K Q, Han T, Liu G, Chen T Y, Wang Y, Yu A Y L, Liu Y L. A geminivirus-based guide RNA delivery system for CRISPR/Cas9 mediated plant genome editing. Sci Rep, 2015,5:14926.
pmid: 26450012
[27] Ali Z, Abul-Faraj A, Li L X, Ghosh N, Piatek M, Mahjoub A, Aouida M, Piatek A, Baltes N J, Voytas D F, Dinesh-Kumar S, Mahfouz M M. Efficient virus-mediated genome editing in plants using the CRISPR/Cas9 system. Mol Plant, 2015,8:1288-1291.
doi: 10.1016/j.molp.2015.02.011 pmid: 25749112
[28] Ali Z, Eid A, Ali S, Mahfouz M M. Pea early-browning virus- mediated genome editing via the CRISPR/Cas9 system in Nicotiana benthamiana and Arabidopsis. Virus Res, 2018,244:333-337.
pmid: 29051052
[29] Cody W B, Scholthof H B, Mirkov T E. Multiplexed gene editing and protein overexpression using aTobacco mosaic virus viral vector. Plant Physiol, 2017,175:23-35.
doi: 10.1104/pp.17.00411 pmid: 28663331
[30] Jiang N, Zhang C, Liu J Y, Guo Z H, Zhang Z Y, Han C G, Wang Y. Development of Beet necrotic yellow vein virus-based vectors for multiple-gene expression and guide RNA delivery in plant genome editing. Plant Biotechnol J, 2019,17:1302-1315.
pmid: 30565826
[31] Gu Z H, Huang C J, Li F F, Zhou X P. A versatile system for functional analysis of genes and microRNAs in cotton. Plant Biotechnol J, 2014,12:638-649.
pmid: 24521483
[32] 雷建峰, 伍娟, 陈晓俊, 於添平, 倪志勇, 李月, 张巨松, 刘晓东. 棉花花粉中高效转录U6启动子的克隆及功能分析. 中国农业科学, 2015,48:3794-3802.
Lei J F, Wu J, Chen X J, Yu T P, Ni Z Y, Li Y, Zhang J S, Liu X D. Cloning and functional analysis of the highly efficient transcription U6 promoter in cotton pollen. Sci Agric Sin, 2015,48:3794-3802 (in Chinese with English abstract).
[33] Zhu S H, Yu X L, Li Y J, Sun Y Q, Zhu Q H, Sun J. Highly efficient targeted gene editing in upland cotton using the CRISPR/Cas9 system. Int J Mol Sci, 2018,19:3000.
[34] Gao W, Long L, Tian X Q, Xu F C, Liu J, Prashant K S, Jose R B, Song C P. Genome editing in cotton with the CRISPR/Cas9 system. Front Plant Sci, 2017,8:1364.
[35] 李妮娜, 丁林云, 张志远, 郭旺珍. 棉花叶肉原生质体分离及目标基因瞬时表达体系的建立. 作物学报, 2014,40:231-239.
Li N N, Ding L Y, Zhang Z Y, Guo W Z. Isolation of mesophyll protoplast and establishment of gene transient expression system in cotton. Acta Agron Sin, 2014,40:231-239 (in Chinese with English abstract).
[1] Peng Jia-Luo, Li Ying, Li Dan-Dan, Yang Jun-Ning, Guo Xue-Feng, Zhang Wen-Jiao, Yu Xiao-Xue, Zhou Ya-Rong, Wang Zhen-Yu, Wang Cai-Xiang, Ma Xiong-Feng, Su Jun-Ji. Identification of class I LBD family members in upland cotton and function and haplotype analyses of GhLBD6 in regulating flowering period [J]. Acta Agronomica Sinica, 2026, 52(6): 1682-1697.
[2] Zhao Jia-Xue, Zhou Long-Hao, Guo Qi-Yuan, Shang Lun-Xiao, Wang Han, Liu Zhi-Tao, Chen Xi, Zhang Xiao-Pei, Song Xian-Liang, Ahmedov Miraziz Baltaevich, Mao Li-Li. Long-term stubble return and subsoiling enhance cotton yields in coastal saline-alkali soils by improving soil conditions and photosynthetic characteristics [J]. Acta Agronomica Sinica, 2026, 52(5): 1548-1560.
[3] Zhang Xi, Wang Guang-En, Li Shao-Qi, Liu Yi, Li Jun-Lan, Qian Yu-Yuan. Transcriptome sequencing-based analysis on the formation mechanism of fiber micronaire differences between two sister lines derived from Gossypium hirsutum-G. barbadense hybrid [J]. Acta Agronomica Sinica, 2026, 52(5): 1442-1458.
[4] Chen Wei, Wei Wan-Juan, Zhao Qi-Bing, Chang Dong-Wei, Yu Ling-Bo, Zhai Peng-Fei, Feng Zhi-Ming, Chen Zong-Xiang, Ren Yang-Tao, Yang Peng, Liu Hai-Lang, Li Zhen-Fu, Yang Yong-Le, Jin Yan-Gang, Zuo Shi-Min. Developing new germplasm of high-quality and early-maturing rice by editing Hd6 via CRISPR/Cas9 [J]. Acta Agronomica Sinica, 2026, 52(4): 1046-1056.
[5] Zhou Qi-Xiang, Zhu Yan, Wang Chu-Bo, Zhu Bo-Lin, Li Jun-Bo, Song Li-Bing. Modeling the effects of climate change on cotton phenology and potential yield in Xinjiang based on the DSSAT model [J]. Acta Agronomica Sinica, 2026, 52(2): 590-602.
[6] LI Yi-Qian, XU Shou-Zhen, LIU Ping, MA Qi, XIE Bin, CHEN Hong. Genome-wide association study of yield components using a 40K SNP array and identification of a stable locus for boll weight in upland cotton (Gossypium hirsutum L.) [J]. Acta Agronomica Sinica, 2025, 51(8): 2128-2138.
[7] GUO Dong-Cai, LYU Tao, CAI Yong-Sheng, MAI WU-LU-DA·AI He-Mai-Ti, CHEN Quan-Jia, QU Yan-Ying, ZHENG Kai. Meta-analysis of QTL and identification of candidate genes for fiber quality in cotton [J]. Acta Agronomica Sinica, 2025, 51(6): 1445-1466.
[8] HUANG Meng-Xin, ZHUANG Ling-Ling, CHENG Pei-Pei, LI Qin, XU Jian-Tang, TAO Ai-Fen, FANG Ping-Ping, QI Jian-Min, ZHANG Li-Wu. Cloning and transcriptional activity analysis of U6 promoter in jute (Corchorus capsularis) [J]. Acta Agronomica Sinica, 2025, 51(5): 1156-1165.
[9] WANG Ya-Wen, QI Zheng-Yang, YOU Jia-Qi, NIE Xin-Hui, CAO Juan, YANG Xi-Yan, TU Li-Li, ZHANG Xian-Long, WANG Mao-Jun. Preparation of cotton 60K functional locus gene chip and its application to genetic research [J]. Acta Agronomica Sinica, 2025, 51(5): 1178-1188.
[10] DING Jun-Feng, XU Ying-Fei, ZHANG Xiang, CHEN Yuan, CHEN De-Hua. Effects of the plant growth regulator IBA on the survival and growth of substrate- grown transplanted cotton seedlings [J]. Acta Agronomica Sinica, 2025, 51(12): 3331-3341.
[11] HALIHASHI Yibati, ZHANG Yan, LI Qing-Jun, XU Xin-Peng, HE Ping. Study on smart fertilizer recommendation methods based on yield response and agronomic efficiency for cotton [J]. Acta Agronomica Sinica, 2025, 51(11): 3052-3064.
[12] ZHAO Hai-Hong, LI Meng-Yuan, LIU Jin-Jing, WANG Yuan-Yuan, DU Lei, WANG Juan, DONG Cheng-Guang, LI Cheng-Qi. Detection of QTNs and QTN-by-environment interactions for plant height in upland cotton (G. hirsutum L.) using the 3VmrMLM method [J]. Acta Agronomica Sinica, 2025, 51(10): 2619-2631.
[13] LI Ya-Wei, XU Ying-Ying, ZUO Chun-Yang, LIU Ruo-Nan, LIANG Ya-Jun, KONG Jie, ZHANG Xian-Long, MIN Ling. Construction of a meiotic progression identification system in cotton and analysis of its response to high-temperature stress [J]. Acta Agronomica Sinica, 2025, 51(10): 2570-2580.
[14] CHEN Jia-Wei, LIN Yan, ZHANG Ming-Xing, ZHOU Shi-Jing, RAO Li-Qun, ZHOU Chi, LI Xin. Effects of Bacillus velezensis YCH92 on the rhizosphere microbial community and yield of cotton [J]. Acta Agronomica Sinica, 2025, 51(10): 2821-2835.
[15] XIE Zhang-Shu, XIE Xue-Fang, TU Xiao-Ju, LIU Ai-Yu, DONG He-Zhong, ZHOU Zhong-Hua. Research progress in phytohormone regulation of square and boll shedding in cotton [J]. Acta Agronomica Sinica, 2025, 51(1): 1-29.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd