Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (9): 1710-1722.doi: 10.3864/j.issn.0578-1752.2022.09.002

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

Establishment of ALSV-Induced Gene Silencing in Chinese Soybean Cultivars

DONG YongXin1(),WEI QiWei1,2(),HONG Hao1,HUANG Ying1,ZHAO YanXiao1,FENG MingFeng1,DOU DaoLong1,*(),XU Yi1,*(),TAO XiaoRong1,*()   

  1. 1College of Plant Protection, Nanjing Agricultural University/Key Laboratory of Soybean Disease and Pest Control of Ministry of Agriculture and Rural Affairs, Nanjing 210095
    2Dazhou Academy of Agricultural Sciences in Sichuan Province, Dazhou 635000, Sichuan
  • Received:2021-11-23 Revised:2022-01-10 Online:2022-05-01 Published:2022-05-19
  • Contact: DaoLong DOU,Yi XU,XiaoRong TAO E-mail:2019102035@njau.edu.cn;675815483@qq.com;taoxiaorong@njau.edu.cn;xuyiqdpd@njau.edu.cn;ddou@njau.edu.cn

RichHTML

95

PDF

643

Abstract:

【Objective】 The establishment of apple latent spherical virus (ALSV)-induced gene silencing on domestic soybean cultivars will offer a simple, time-saving and operable system for gene function and genetic breeding of soybean cultivars in China.【Method】The infectious clones of ALSV which was used to infect plant through Agro-infiltration was constructed. A 327 bp cDNA fragment of phytoene desaturase (GmPDS) cDNA was amplified from soybean cultivar Williams 82 and inserted into the pALSV2 vector. The pALSV1 and pALSV2-GmPDS were used to co-infect Nicotiana benthamiana plant through Agro-infiltration, and the ALSV virus particle was purified from systemic infected leaves of N. benthamiana plants at 17 days post infiltration (dpi). The purified virions were mechanically inoculated onto the first-round true leaves of soybean and photobleaching phenotype on the upper non-inoculated leaves was monitored from 10 to 30 dpi. Using the empty ALSV vector as a control, the gene expression levels of ALSV coat protein (CP) gene and GmPDS in ALSV:GmPDS infected soybean were examined by RT-PCR and quantitative RT-PCR analysis, respectively. 【Result】At 20 dpi, soybean cultivar Nannong 1138-2 inoculated with ALSV:GmPDS showed photobleaching phenotype in the systemic infected leaves, however Williams 82 did not show gene silencing phenotype of PDS in the systemic infected leaves. The results of qRT-PCR confirmed that the RNA expression level of GmPDS in Nannong 1138-2 was significantly reduced but ALSV:GmPDS was unable to silence PDS in Williams 82. By using the same method, the gene silencing efficiency of other 9 domestic soybean cultivars mediated by ALSV:GmPDS was examined. Soybean cultivars including Nannong 47, Andou 203, Xiangdou 4, Zhonghuang 13, Shanning 29 and Qihuang 34 all displayed photobleaching phenotype in plant systemic infected leaves after inoculation with ALSV:GmPDS, while GmPDS was not silenced in Hedou 12, Zhonghuang 311 and Shanning 16.【Conclusion】In this study, an Agrobacterium-mediated ALSV viral VIGS vector was constructed. ALSV virions was propagated and purified from N. benthamiana plant. The purified virions were rub inoculated onto the first-round true leaves of soybean. Finally, an efficient gene silencing system was established on a number of domestic soybean cultivars in China.

Key words: virus-induced gene silencing (VIGS), soybean, apple latent spherical virus, phytoene desaturase (PDS)

Table 1

Primers used in this study"

引物名称
<BOLD>P</BOLD>rimer name		 序列
Sequence of the primers (5′-3′)		 用途
Purpose
GmPDSF CTGATTTCACACTCGAGCCCGCTGACATGTCAGTAACTTGC GmPDS扩增
Amplification of GmPDS
GmPDSR CCTTCTAGCAGGGATCCCCCCATTGAAGCTAAATATTTTTG
GmActinF GATACTAGACTATCGTTCTCCTAACTCG 大豆内参引物（RT-PCR）
Internal control primers (RT-PCR)
GmActinR GCATGAAAATCAAGGTGGTTGC
qGmActinF TAGAGCCACCAATCCAGACAC 大豆内参引物（qRT-PCR）
Internal control primers (qRT-PCR)
qGmActinR TCGTATGAGCAAGGAAATTGG
qGmPDSF CCTGAACGGGTAACTGATGAGGTG GmPDS沉默验证
Verification of the silence efficiency of GmPDS
qGmPDSR CGGCATACAAAGTCTTTCGGGTG
ALSV2F CTTGTGTATGATGAACTCATGCAATTG ALSV CP引物
ALSV CP primers
ALSV2R GAAGCTTAGGAAAGCGAGTGCG
1F GCATAGATCTGACCCAAATCTGC 验证目的片段
Verification of the target fragment
1R CTTTGTGAAATCAGGGCCCTGAC
NbEF1αF TGCTGCAACAAGATGGATGC 本氏烟内参引物
Internal control primers for Nicotiana benthamiana
NbEF1αR CCAGAGATGGGGACAAAGGG

Fig. 1

Construction of pALSV2-GmPDS silencing vector A: Schematic diagram of pALSV2-GmPDS construction. P35S: Enhanced CaMV 35S promoter; Tnos: Nopaline synthase terminator; PRO-co: Protease cofactor; HEL: NTP-binding helicase; C-PRO: Cysteine protease; POL: RNA polymerase; MP: 42K movement protein; Vp25, Vp20, and Vp24: Capsid proteins. B: Sequence alignment of GmPDS cDNA amplified regions"

Fig. 2

The procedure of ALSV virus particle extraction and enrichment from infected Nicotiana benthamiana plants"

Fig. 3

Phenotypes and RT-PCR detection of ALSV in systemic leaves of Nicotiana benthamiana by Agroinfitration A: Agrobacterium-mediated ALSV infection of N. benthamiana plant. Phenotypes of N. benthamiana at 17 d after infiltrated with 1﹕1﹕1 mixture of agrobacteria containing pALSV1, pALSV2-derivative vectors or P19. MOCK: Infiltrated with empty vector; P19: Gene silencing suppressor; B: The content of ALSV CP in systemic leaves was detected by RT-PCR; C: Agrobacterium-mediated ALSV﹕GmPDS infection of N. benthamiana plant and effect of P19 on ALSV infection; CH42: Chlorophyll synthesis related genes; D: The ALSV CP and inserted CH42 or GmPDS sequences in systemic leaves of plants in panel C was detected by RT-PCR"

Table 2

Gene silencing efficiency in eleven soybean cultivars using ALSV induced VIGS"

品种
Cultivar		 ALSV系统侵染
Systemic infection of
ALSV		 系统叶变白
<BOLD>P</BOLD>hotobleaching in systemic leaves		 白化植物数/接种植物数
No. of plants showing photo-bleaching/No. of inoculated plants
南农1138-2 Nannong 1138-2 √ √ 6/6
威廉姆斯82 Williams 82 √ × 0/6
齐黄34 Qihuang 34 √ √ 3/8
安豆203 Andou 203 √ √ 3/8
南农47 Nannong 47 √ √ 3/8
祥斗4号 Xiangdou 4 √ √ 4/8
中黄13 Zhonghuang 13 √ √ 4/6
山宁29 Shanning 29 √ √ 4/6
山宁16 Shanning 16 √ × 0/6
菏豆12 Hedou 12 √ × 0/6
中黄311 Zhonghuang 311 √ × 0/6

Fig. 4

Phenotypes and qRT-PCR detections of Nannong 1138-2 and Williams 82 mechanically inoculated with ALSV:GmPDS virus particle A: Phenotypes of Nannong 1138-2 mechanically inoculated with ALSV virus particles at 20 days post inoculation (dpi); B: Phenotypes of Nannong 1138-2 inoculated with ALSV:GmPDS virus particles at 20 dpi; C: The phenotypes of systemic leaves of Nannong 1138-2 soybean cultivar treated with ALSV and ALSV:GmPDS virus particles from Figure A and B; left leaves were inoculated with ALSV virus particles, and right leaves were inoculated with ALSV:GmPDS virus particles; D: qRT-PCR was used to quantify GmPDS silencing efficiency in systemic leaves of Nannong 1138-2 soybean cultivar treated with ALSV and ALSV:GmPDS virus particles (n=3, **: P<0.01); E: Phenotypes of Williams 82 inoculated with ALSV:GmPDS virus particles at 20 dpi; F: qRT-PCR was used to detect relative RNA expression of GmPDS in systemic leaves of Williams 82 treated with ALSV and ALSV:GmPDS virus particles (n=3, ns: P>0.05)"

Fig. 5

Detection of ALSV accumulation in systemic leaves of Williams 82 +: Using cDNA from ALSV systemically infected leaves of Nannong 1138-2 showing photobleaching phenotype as a positive control; —: cDNA from healthy soybean as a negative control"

Fig. 6

Determination of virus induced gene silencing efficiency in nine different domestic soybean cultivars using ALSV:GmPDS Phenotypes of nine different soybean cultivars mechanically inoculated with ALSV:GmPDS virus particles at 20 dpi; ALSV:GmPDS treated Qihuang 34, Nannong 47, Xiangdou 4, Zhonghuang 13, Shanning 29 and Andou 203 all showed leave photobleaching phenotype; Shanning 16, Hedou 12 and Zhonghuang 311 did not show photobleaching phenotype"

Fig. 7

Detection of ALSV accumulation in systemic leaves of different soybean cultivars A: The accumulation of CP could be detected in the systemic leaves of Qihuang 34 inoculated with ALSV virus particles, and CP gene could be detected in three out of eight samples inoculated with ALSV:GmPDS; B: CP could not be detect or detected only with a very low accumulation in Hedou 12 inoculated with ALSV and ALSV:GmPDS, respectively; C: CP could only be detected with a very low accumulation in Zhonghuang 311 inoculated with ALSV and ALSV:GmPDS; D: CP could only be detected with a very low accumulation in Shanning 16 inoculated with ALSV and ALSV:GmPDS, respectively. +: Using cDNA from ALSV systemically infected leaves of Nannong 1138-2 showing photobleaching phenotype as a positive control. —: cDNA from healthy soybean as a negative control"

[1] 吴汶珊. 中国大豆对外贸易的发展现状及对策. 大陆桥视野, 2021(6): 88-89.
WU W S. Development status and countermeasures of Chinese soybean foreign trade. New Silk Road Horizon, 2021(6): 88-89. (in Chinese)
[2] 吴曰程, 王玉斌. 中国转基因大豆进口及其影响分析. 大豆科学, 2019, 38(4): 635-643.
WU Y C, WANG Y B. Import of transgenic soybean in China and its impact analysis. Soybean Science, 2019, 38(4): 635-643. (in Chinese)
[3] 刘晓彬, 刘娜, 李福宽, 吴立柱, 张洁, 王冬梅. TRV介导的大豆基因瞬时沉默体系的建立. 中国农业科学, 2015, 48(12): 2479-2486.
LIU X B, LIU N, LI F K, WU L Z, ZHANG J, WANG D M. Establishment of TRV-mediated transient gene-silencing system in soybean. Scientia Agricultura Sinica, 2015, 48(12): 2479-2486. (in Chinese)
[4] 王爽, 郭兵福, 郭勇, 张丽娟, 金龙国, 杨慧, 邱丽娟. 病毒诱导的基因沉默(VIGS)技术及其在大豆基因功能研究和育种中的应用潜力. 大豆科学, 2016, 35(4): 536-540.
WANG S, GUO B F, GUO Y, ZHANG L J, JIN L G, YANG H, QIU L J. Virus-induced gene silencing and its usage in soybean functional genomics. Soybean Science, 2016, 35(4): 536-540. (in Chinese)
[5] 李敏, 关博文, 杨学, 陈庆山, 张睿, 武小霞, 金慧, 吴玉娥. 大豆种质资源遗传多样性分析. 农业科技通讯, 2021(11): 4-8.
LI M, GUAN B W, YANG X, CHEN Q S, ZHANG R, WU X X, JIN H, WU Y E. Genetic diversity analysis of soybean germplasm resources. Bulletin of Agricultural Science and Technology, 2021(11): 4-8. (in Chinese)
[6] LANGE M, YELLINA A L, ORASHAKOVA S, BECKER A. Virus-induced gene silencing (VIGS) in plants: An overview of target species and the virus-derived vector systems. Methods in Molecular Biology, 2013, 975: 1-14.
[7] RAMEGOWDA V, MYSORE K S, SENTHIL-KUMAR M. Virus-induced gene silencing is a versatile tool for unraveling the functional relevance of multiple abiotic-stress-responsive genes in crop plants. Frontiers of Plant Science, 2014, 5: 323.
[8] BURCH-SMITH T M, ANDERSON J C, MARTIN G B, DINESH-KUMAR S P. Application and advantages of virus-induced gene silencing for gene function studies in plants. The Plant Journal, 2004, 39: 734-746.
doi: 10.1111/j.1365-313X.2004.02158.x
[9] KANAZAWA A, INABA J I, SHIMURA H, OTAGAKI S, TSUKAHARA S, MATSUZAWA A, KIM B M, GOTO K, MASUTA C. Virus-mediated efficient induction of epigenetic modifications of endogenous genes with phenotypic changes in plants. The Plant Journal, 2011, 65(1): 156-168.
doi: 10.1111/j.1365-313X.2010.04401.x
[10] LU R, MARTIN-HERNANDEZ A M, PEART J R, MALCUIT I, BAULCOMBE D C. Virus-induced gene silencing in plants. Methods, 2003, 30(4): 296-303.
doi: 10.1016/S1046-2023(03)00037-9
[11] ZHANG C, GHABRIAL S A. Development of bean pod mottle virus-based vectors for stable protein expression and sequence- specific virus-induced gene silencing in soybean. Virology, 2006, 344(2): 401-411.
doi: 10.1016/j.virol.2005.08.046
[12] KACHROO A, GHABRIAL S. Virus-induced gene silencing in soybean. Methods in Molecular Biology, 2012, 894: 287-297.
[13] IGARASHI A, YAMAGATA K, SUGAI T, TAKAHASHI Y, SUGAWARA E, TAMURA A, YAEGASHI H, YAMAGISHI N, TAKAHASHI T, ISOGAI M, TAKAHASHI H, YOSHIKAWA N. Apple latent spherical virus vectors for reliable and effective virus-induced gene silencing among a broad range of plants including tobacco, tomato, Arabidopsis thaliana, cucurbits, and legumes. Virology, 2009, 386(2): 407-416.
doi: 10.1016/j.virol.2009.01.039
[14] GEDLING C R, ALI E M, GUNADI A, FINER J J, XIE K, LIU Y, YOSHIKAWA N, QU F DORRANCE A E. Improved apple latent spherical virus-induced gene silencing in multiple soybean genotypes through direct inoculation of agro-infiltrated Nicotiana benthamiana extract. Plant Methods, 2018, 14: 19.
doi: 10.1186/s13007-018-0286-7
[15] LI C, YOSHIKAWA N, TAKAHASHI T, ITO T, YOSHIDA K, KOGANEZAWA H. Nucleotide sequence and genome organization of apple latent spherical virus: A new virus classified into the family comoviridae. Journal of General Virology, 2000, 81(2): 541-547.
[16] YAMAGISHI N, SASAKI S, YAMAGATA K, KOMORI S, NAGASE M, WADA M, YAMAMOTO T, YOSHIKAWA N. Promotion of flowering and reduction of a generation time in apple seedlings by ectopical expression of the Arabidopsis thaliana FT gene using the apple latent spherical virus vector. Plant Molecular Biology, 2011, 75(1/2): 193-204.
doi: 10.1007/s11103-010-9718-0
[17] HUANG Y, HONG H, XU M, YAN J, DAI J, WU J, FENG Z, ZHU M, ZHANG Z, YUAN X, DING X, TAO X. Developmentally regulated Arabidopsis thaliana susceptibility to tomato spotted wilt virus infection. Molecular Plant Pathology, 2020, 21(7): 985-998.
doi: 10.1111/mpp.12944
[18] KON T, YOSHIKAWA N. Induction and maintenance of DNA methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing. Frontiers in Microbiology, 2014, 5: 595.
[19] LI C, HIRANO H, KASAJIMA I, YAMAGISHI N, YOSHIKAWA N. Virus-induced gene silencing in chili pepper by apple latent spherical virus vector. Journal of Virological Methods, 2019, 273: 113711.
doi: 10.1016/j.jviromet.2019.113711
[20] OGATA T, TOYOSHIMA M, YAMAMIZO-ODA C, KOBAYASHI Y, FUJII K, TANAKA K, TANAKA T, MIZUKOSHI H, YASUI Y, NAGATOSHI Y, YOSHIKAWA N, FUJITA Y. Virus-mediated transient expression techniques enable functional genomics studies and modulations of betalain biosynthesis and plant height in Quinoa. Frontiers of Plant Science, 2021, 12: 643499.
doi: 10.3389/fpls.2021.643499
[21] YAMAGISHI N, YOSHIKAWA N. Highly efficient virus-induced gene silencing in apple and soybean by apple latent spherical virus vector and biolistic inoculation. Methods in Molecular Biology, 2013, 975: 167-181.
[22] 高鹏飞, 席飞虎, 张泽宇, 胡凯强, 陈凯, 魏文桃, 丁家治, 顾连峰. 植物VIGS技术及其在林业科学中的研究进展. 生物技术通报, 2021, 37(5): 141-153.
doi: 10.13560/j.cnki.biotech.bull.1985.2020-1452
GAO P F, XI F H, ZHANG Z Y, HU K Q, CHEN K, WEI W T, DING J Z, GU L F. Research progress of plant VIGS technology and its application in forestry science. Biotechnology Bulletin, 2021, 37(5): 141-153. (in Chinese)
doi: 10.13560/j.cnki.biotech.bull.1985.2020-1452
[23] WERNER R C, DUGÉ DE BERNONVILLE T, GLÉVAREC G, LANOUE A, OUDIN A, PICHON O, ST-PIERRE B, COURDAVAULT V, BESSEAU S. ALSV-based virus-induced gene silencing in apple tree (Malus×domestica L.). Methods in Molecular Biology, 2020, 2172: 183-197.
[24] 周彦. 果树病毒载体研究进展. 中国农业科学, 2014, 47(6): 1119-1127.
ZHOU Y. Progresses in study of virus-based vectors of fruit trees. Scientia Agricultura Sinica, 2014, 47(6): 1119-1127. (in Chinese)
[25] YAEGASHI H, YAMATSUTA T, TAKAHASHI T, LI C, ISOGAI M, KOBORI T, OHKI S, YOSHIKAWA N. Characterization of virus-induced gene silencing in tobacco plants infected with apple latent spherical virus. Archives of Virology, 2007, 152(10): 1839-1849.
doi: 10.1007/s00705-007-1011-4
[26] BURCH-SMITH T M, SCHIFF M, LIU Y, DINESH-KUMAR S P. Efficient virus-induced gene silencing in Arabidopsis. Plant Physiology, 2006, 142(1): 21-27.
doi: 10.1104/pp.106.084624
[27] XU H Y, ZHANG C, LI Z C, WANG Z R, JIANG X X, SHI Y F, TIAN S N, BRAUN E, MEI Y, QIU W L, LI S, WANG B, XU J, NAVARRE D, REN D, CHENG N, NAKATA P A, GRAHAM M A, WHITHAM S A, LIU J Z. The MAPK kinase kinase GmMEKK1 regulates cell death and defense responses. Plant Physiology, 2018, 178(2): 907-922.
doi: 10.1104/pp.18.00903
[28] TIAN S N, LIU D D, ZHONG C L, XU H Y, YANG S, FANG Y, RAN J, LIU J Z. Silencing GmFLS2 enhances the susceptibility of soybean to bacterial pathogen through attenuating the activation of GmMAPK signaling pathway. Plant Science, 2020, 292: 110386.
doi: 10.1016/j.plantsci.2019.110386
[29] LI C, LI K, ZHENG M, LIU X, DING X, GAI J, YANG S. Gm6PGDH1, a cytosolic 6-phosphogluconate dehydrogenase, enhanced tolerance to phosphate starvation by improving root system development and modifying the antioxidant system in soybean. Frontiers in Plant Science, 2021, 12: 704983.
doi: 10.3389/fpls.2021.704983
[30] KIM K H, LIM S, KANG Y J, YOON M Y, NAM M, JUN T H, SEO M J, BAEK S B, LEE J H, MOON J K, LEE S H, LEE S H, LIM H S, MOON J S, PARK C H. Optimization of a virus-induced gene silencing system with soybean yellow common mosaic virus for gene function studies in soybeans. Plant Pathology Journal, 2016, 32(2): 112-122.
[1] LI YongJuan, ZHANG YueTong, WANG YiBo, ZHAO ChangJiang, SONG Jie, CHEN XueLi, YAO Qin. Effects of Biochar Application on the Abundance and Community Composition of Nitrogen-Fixing Microbial nifH Gene in Soybean Rotation and Continuous Cropping Systems [J]. Scientia Agricultura Sinica, 2026, 59(6): 1272-1285.
[2] LIU FangDong, SUN Lei, WANG WuBin, ZHAO JinMing, GAI JunYi. Changes of Cropping System and Suggestions on Ecological Cultivation Regions of Soybeans in China [J]. Scientia Agricultura Sinica, 2026, 59(3): 486-498.
[3] CAI TingYang, ZHU YuPeng, LI RuiDong, WU ZongSheng, XU YiFan, SONG WenWen, XU CaiLong, WU CunXiang. Effects of Leaf-Cutting at Seedling Stage on Photosynthetic Characteristics, Pod Distribution and Yield Formation in Soybean in the Huang-Huai-Hai Region [J]. Scientia Agricultura Sinica, 2026, 59(2): 292-304.
[4] WU Qiong, XIE XiangTing, WANG Lei, MOU Yong, LI JinWei. Development and Validation of Event-Specific PCR Method for the Quantification of Genetically Modified Soybean DBN8205 [J]. Scientia Agricultura Sinica, 2026, 59(1): 29-40.
[5] LIU LuPing, HU XueJie, QI Jin, CHEN Qiang, LIU Zhi, ZHAO TianTian, SHI XiaoLei, LIU BingQiang, MENG QingMin, ZHANG MengChen, HAN TianFu, YANG ChunYan. Cloning of the Promoters and Analysis of Expression Patterns of Maturity Genes E1 and E2 in Soybean [J]. Scientia Agricultura Sinica, 2025, 58(5): 840-850.
[6] ZHENG Yu, CHEN Yi, TI JinSong, SHI LongFei, XU XiaoBo, LI YuLin, GUO Rui. Evaluation of Carbon Footprint and Economic Benefit of Different Tobacco Rotation Patterns [J]. Scientia Agricultura Sinica, 2025, 58(4): 733-747.
[7] ZHANG Qi, XUE FuZhen, YANG XiuJie, JIANG SuYang, HUANG XueJuan, MA JiaYi, ZHANG ZheWen, XU JieFei. Study on the Function of Soybean Nicotinamide Enzyme GmNIC1 Gene Under Saline Alkali Stress [J]. Scientia Agricultura Sinica, 2025, 58(24): 5128-5142.
[8] MA HeXiao, GE GuoLong, ZHANG XiangQian, LU ZhanYuan, WANG ManXiu, RONG MeiRen, SHI JingJing, ZHANG DeJian, SUN XuePing. Effects of Different Crop Rotation Systems on Soil Readily Oxidized Organic Carbon and Carbon Pool Activity Differences [J]. Scientia Agricultura Sinica, 2025, 58(24): 5201-5215.
[9] GAO ChunHua, ZHAO HaiJun, ZHAO FengTao, KONG WeiLin, JU FeiYan, LI ZongXin, SHI DeYang, LIU Ping. Effect of Growth Regulators on the Stem Characteristics and Yield of Summer Maize in Maize-Soybean Strip Intercropping [J]. Scientia Agricultura Sinica, 2025, 58(23): 4920-4935.
[10] YANG ShuQi, ZHAO YingXing, QIAN Xin, ZHANG XuePeng, MENG WeiWei, SUI Peng, LI ZongXin, CHEN YuanQuan. Comprehensive Evaluation of the Maize-Soybean Intercropping Pattern in the Huang-Huai Region [J]. Scientia Agricultura Sinica, 2025, 58(23): 4936-4951.
[11] FANG Jian, QIN ZhaoJi, YU YuanYuan, YU NingNing, ZHAO Bin, LIU Peng, REN BaiZhao, ZHANG JiWang. Impacts of Varying Row Ratio Arrangements on Plant Performance, Stand Yield, and Comprehensive Benefits in Soybean-Maize Strip intercropping [J]. Scientia Agricultura Sinica, 2025, 58(23): 4841-4857.
[12] SONG XuHui, ZHAO XueYing, ZHAO Bin, REN BaiZhao, ZHANG JiWang, LIU Peng, REN Hao. Effects of Row Ratio Allocation on Light Distribution and Photosynthetic Production Capacity of Maize-Soybean Strip Intercropping [J]. Scientia Agricultura Sinica, 2025, 58(23): 4858-4871.
[13] SHI DeYang, GAO ChunHua, LI YanHong, ZHAO HaiJun, XIA DeJun. Effects of Row Spacing Configuration on the Canopy Characteristics and Grain Yield of the Intercropping Maize [J]. Scientia Agricultura Sinica, 2025, 58(23): 4872-4885.
[14] ZHANG MengYu, HE ZaiJu, WANG XingXing, REN Hao, REN BaiZhao, LIU Peng, ZHANG JiWang, ZHAO Bin. The Influences of Different Plant Height Combinations of Maize Varieties on Light Distribution in the Canopy and the Photosynthetic Characteristics of Maize Under Maize-Soybean Strip Intercropping Pattern [J]. Scientia Agricultura Sinica, 2025, 58(23): 4886-4904.
[15] KONG WeiLin, GAO ChunHua, ZHAO FengTao, JU FeiYan, LI ZongXin, ZHAO HaiJun, LIU Ping. Effects of Nitrogen Application Rate Combined with Drip Irrigation Amount After Sowing on Yield, Economic Benefit, Water Use Characteristics of Maize-Soybean Strip Intercropping Planting System [J]. Scientia Agricultura Sinica, 2025, 58(23): 4905-4919.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd