小麦Glu-3位点基因拷贝数的变异分析

doi:10.3864/j.issn.0578-1752.2021.06.002

Abstract

Abstract:

【Objective】The variations in numbers of copies of a gene is a common and important gene structure variation, which often effects individual phenotype. Low-molecular-weight glutenin subunit (LMW-GS) located at the Glu-3 loci, is an essential part of storage protein in wheat. As a hetero-hexaploid, the huge and complex wheat genome makes it difficult to detect numbers of gene copies by classical methods. A limited information is available about CNV analysis of LWM-GS. To screen reliable and stable internal reference genes and systems for complex genome and explore the CNV determination technology suitable in different wheat varieties, the techniques used for detection of gene copy numbers, based on droplet digital PCR (ddPCR) which improve the detection flux of target gene and determine the particular numbers of gene copies of Glu-3 loci. 【Method】In this paper, Acc1 was used as the internal reference gene. The internal reference primers and probes were designed by using the corresponding gene sequence. In order to analyze stability of numbers of copies of Acc1 in 12 common wheat varieties, we used qualitative and quantitative PCR. The genomic DNA of variety Gaoyou2018 with five different dilutions was used as template, and qRT-PCR was used to analyze the repeatability and accuracy of Acc1 internal reference system (primers and probes).The corresponding specific primers and probes were designed according to the LMW-GS gene sequence of Glu-A3 loci. Two different methods, qRT-PCR and ddPCR, were used to detect the numbers of copies of Glu-A3 loci in eight wheat varieties, in order to determine which method is more suitable for high-throughput detection of Glu-3 loci gene. The specific primers and probes were also designed according to the sequences of LMW-GS gene at Glu-B3 and Glu-D3 loci. The numbers of copies of LMW-GS gene at Glu-A3, Glu-B3 and Glu-D3 loci of 231 varieties were determined and analyzed by ddPCR. 【Result】The results showed that the gene copy numbers of Acc1 was consistent among varieties and different DNA concentrations of the same variety. The coefficient of variation (CV) between repeats was 0.07%-0.77%. It is also indicated from results that the Acc1 internal reference system constructed in this paper has good stability and repeatability. The results of qRT-PCR and ddPCR were consistent in detecting the numbers of copies of LMW-GS gene at Glu-A3 loci in 8 wheat varieties, which were 3, 5, 3, 4, 3, 3, 3 and 3, respectively. However, the CV among repeats detected by ddPCR was 0.30%-1.67%, which much lower than that by qRT-PCR. It showed that using ddPCR method to detect the gene copy numbers of Glu-3 loci gene is more stable and reliable. The numbers of copies of LMW-GS gene were 4 at Glu-A3, Glu-B3 and Glu-D3 having a frequency of 95%, 32.03% 28.57% respectively in 231 wheat varieties. The total variation range in numbers of copies of Glu-3 was 10-21, and the CV was 16.12%.【Conclusion】With good stability and repeatability, Acc1 could be used as internal reference gene for numbers of gene copies detection in wheat. Both qRT-PCR and ddPCR could be used in gene copy numbers detection with wheat genes, but the ddPCR is more simple, stable, reliable and has high detection flux.

Key words: wheat, low-molecular-weight glutenin subunit (LMW-GS), droplet digital PCR (ddPCR), copy number variation (CNV)

Can CHEN,NanNan HAN,Yang LIU,XiaoWei SHI,HongQi SI,ChuanXi MA. Analysis of Copy Number Variation of Glu-3 Locus in Common Wheat[J].Scientia Agricultura Sinica, 2021, 54(6): 1092-1103.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2021.06.002

https://www.chinaagrisci.com/EN/Y2021/V54/I6/1092

Figures/Tables 10

Table 1

Fig. 1

Fig. 2

Table 2

Fig. 3

Fig. 4

Table 3

Fig. 5

Fig. 6

Fig. 7

References 51

[1]	赵献林, 夏先春, 刘丽, 何中虎, 孙其信. 小麦低分子量麦谷蛋白亚基及其编码基因研究进展. 中国农业科学, 2007,40(3):440-446.
	ZHAO X L, XIA X C, LIU L, HE Z H, SUN Q X. Review on low molecular weight glutenin subunits and their coding genes. Scientia Agricultura Sinica, 2007,40(3):440-446. (in Chinese)
[2]	XIANG L, HUANG L, GONG F Y, LIU J, WANG Y F, JIN Y R, HE Y, HE J S, JIANG Q T, ZHENG Y L, LIU D C, WU B H. Enriching LMW-GS alleles and strengthening gluten properties of common wheat through wide hybridization with wild emmer. 3 Biotech, 2019,9(10):355.
[3]	HAZARD B, TRAFFORD K, LOVEGROVE A, GRIFFITHS S, UAUY C, SHEWRY P. Strategies to improve wheat for human health. Nature Food, 2020,1(8):475-480.
[4]	GUPTA R B, SHEPHERD K W. Two-step one-dimensional SDS-PAGE analysis of LMW subunits of glutelin. Theoretical and Applied Genetics, 1990,80(1):65-74.
[5]	SINGH N K, SHEPHERD K W. Linkage mapping of genes controlling endosperm storage proteins in wheat. Theoretical and Applied Genetics, 1988,75(4):628-641.
[6]	D'OVIDIO R, MASCI S. The low-molecular-weight glutenin subunits of wheat gluten. Journal of Cereal Science, 2004,39(3):321-339.
[7]	CHO K, JO Y M, LIM S H, KIM J Y, HAN O, LEE J Y. Overexpressing wheat low-molecular-weight glutenin subunits in rice (Oryza sativa L. japonica cv. Koami) seeds. 3 Biotech, 2019,9(2):1-8.
[8]	RAI A, SINGH A M, GANJEWALA D, KUMAR R R, AHLAWAT A K, SINGH S K, SHARMA P, JAIN N. Rheological evaluations and molecular marker analysis of cultivated bread wheat varieties of India. Journal of Food Science and Technology, 2019,56(4):1696-1707.
[9]	BEOM H R, KIM J S, JANG Y R, LIM S H, KIM C K, LEE C K, LEE J Y. Proteomic analysis of low-molecular-weight glutenin subunits and relationship with their genes in a common wheat variety. 3 Biotech, 2018,8(1):56.
[10]	SHAW-SMITH C, REDON R, RICKMAN L. Microarray based comparative genomic hybridisation (array-CGH) detects submicroscopic chromosomal deletions and duplications in patients with learning disability/mental retardation and dysmorphic features. Journal of Medical Genetics, 2004,41(4):241-248.
[11]	MARON L G, GUIMARAES C T, KIRST M, ALBERT P S, BIRCHLER J A, BRADBYRY P J, BUCKLER E S, COLUCCIO A E, DANILOVA T V, KUDRNA D, MAGALHAES J V, PINEROS M A, SCHATZ M C, WING R A, KOCHIAN L. Aluminum tolerance in maize is associated with higher MATE1 gene copy number. Proceedings of the National Academy of Sciences of the United States of America, 2013,110(13):5241-5246.
[12]	VOGELSTEIN B, KINZLER K W. Digital PCR. Proceedings of the National Academy of Sciences of the United States of America, 1999,96(16):9236-9241.
[13]	SYKES P J, NEOH S H, BRISCO M J. Quantitation of targets for PCR by use of limiting dilution. Biotechniques, 1992,13(3):444-449.
[14]	YANG Q, XI J, CHEN X X, HU S H, CHEN N, QIAO S L, WAN S G, BAO D K. The development of a sensitive droplet digital PCR for quantitative detection of porcine reproductive and respiratory syndrome virus. International Journal of Biological Macromolecules, 2017,104(Part A):1223-1228.
[15]	LIN Q, FU X Z, LIU L H, LIANG H R, NIU Y J, WEN Y Y, HUANG Z B, LI N Q. Development and application of a sensitive droplet digital PCR (ddPCR) for the detection of infectious spleen and kidney necrosis virus. Aquaculture, 2020,529:735697.
[16]	ANTKOWIAK M, NOWACKA-WOSZUK J, SZCZERBAL I, SWITONSKI M, SZYDLOWSKI M. AMY2B gene copy-number variation studied by droplet digital PCR (ddPCR) in three canids: Red fox, arctic fox, and Chinese raccoon dog. Folia Biologica, 2020,68(2):51-55.
[17]	KONISHI T, SHINOHARA K, YAMADA K, SASAKI Y. Acetyl-CoA carboxylase in higher plants: Most plants other than gramineae have both the prokaryotic and the eukaryotic forms of this enzyme. Plant and Cell Physiology, 1996,37(2):117-122.
[18]	CHEN Q, SONG J, DU W P, XU L Y, JIANG Y, ZHANG J, ZHANG M, YU G R. Phylogenetic analyses of four Chinese endemic wheat landraces based on two single copy genes. Cereal Research Communications, 2018,46(2):191-200.
[19]	雷映霞. 鹅观草属及其近缘属物种的分子系统与进化研究[D]. 雅安: 四川农业大学, 2018.
	LEI Y X. Phylogenetic and evolution analysis of Roegneria and its related genera (Triticeae Poaceae)[D]. Yaan: Sichuan Agricultural University, 2018. (in Chinese)
[20]	GORNICKI P, PODKOWINSKI J, SCAPPINO L A, DIMAIO J, WARD E, HASELKORN R. Wheat acetyl-CoA carboxylase: cDNA and protein structure. Proceedings of the National Academy of Sciences of the United States of America, 1994,91(15):6860-6864.
[21]	PODKOWINSKI J, SROGA G E, HASELKORN R, GORNICKI P. Structure of a gene encoding a cytosolic acetyl-CoA carboxylase of hexaploid wheat. Proceedings of the National Academy of Sciences of the United States of America, 1996,93(5):1870-1874.
[22]	GORNICKI P, FARIS J, KING I, PODKOWINSKI J, GILL B, HASELKORN R. Plastid-localized acetyl-CoA carboxylase of bread wheat is encoded by a single gene on each of the three ancestral chromosome sets. Proceedings of the National Academy of Sciences of the United States of America, 1997,94(25):14179-14184.
[23]	缪青梅, 汪小福, 陈笑芸, 彭城, 徐晓丽, 魏巍, 徐俊锋. 基于双重微滴数字PCR精准定量转基因水稻G6H1的方法研究. 农业生物技术学报, 2019,27(1):159-169.
	MIAO Q M, WANG X F, CHEN X Y, PENG C, XU X L, WEI W, XU J F. Studies on accurate quantification of genetically modified rice (Oryza sative) G6H1 based on duplex droplet digital PCR. Journal of Agricultural Biotechnology, 2019,27(1):159-169. (in Chinese)
[24]	蔡教英, 姚丽锋, 王小玉, 游淑珠, 丁琦. 基于双重微滴式数字PCR对转基因油菜RF1品系的定量方法. 现代食品科技, 2018,34(6):282-287.
	CAI J Y, YAO L F, WANG X Y, YOU S Z, DING Q. Quantitative analysis of genetically modified rapeseed of RF1 by duplex droplet digital polymerase chain reaction (duplex-ddPCR). Modern Food Science and Technology, 2018,34(6):282-287. (in Chinese)
[25]	WENG H B, PAN A H, YANG L T, ZHANG C M, LIU Z L, ZHANG D B. Estimating number of transgene copies in transgenic rapeseed by real-time PCR assay with HMG I/Y as an endogenous reference gene. Plant Molecular Biology Reporter, 2004,22(3):289-300.
[26]	CAVIGLIA G P, ABATE M L, TANDOI F, CIANCIO A, AMOROSO A, SALIZZONI M, SARACCO G M, RIZZETTO M, ROMAGNOLI R, SMEDILE A. Quantitation of HBV cccDNA in anti-HBc-positive liver donors by droplet digital PCR: A new tool to detect occult infection. Journal of Hepatology, 2018,69(2):301-307.
[27]	DYAVAR S R, YE Z, BYRAREDDY S N, SCARSI K K, WINCHESTER L C, WEINHOLD J A, FLETCHER C V, PODANY A T. Normalization of cell associated antiretroviral drug concentrations with a novel RPP30 droplet digital PCR assay. Scientific Reports, 2018,8(1):3626.
[28]	ELMAHALAWY S T, HALVARSSON P, SKARIN M, HÖGLUND J. Genetic variants in dyf-7 validated by droplet digital PCR are not drivers for ivermectin resistance in Haemonchus contortus. International Journal for Parasitology: Drugs and Drug Resistance, 2018,8(2):278-286.
[29]	YU R L, XUAN W J, ZHOU L, LUO Y, LIU X Y, XIONG P W, REN X Y. Detection of HER2 amplification in formalin-fixed paraffin- embedded breast carcinoma tissue with digital PCR using two TFF3 sequences as internal reference. Experimental and Molecular Pathology, 2018,104(3):235-238.
[30]	琚鹏举, 孙黛珍, 宁蕾, 葛林豪, 许成杰, 史华伟, 梁凯歌, 马亮, 刘陶然, 陈明. 采用优化的数字PCR方法分析转基因小麦外源基因拷贝数. 中国农业科学, 2020,53(10):1931-1939.
	JU P J, SUN D Z, NING L, GE L H, XU C J, SHI H W, LIANG K G, MA L, LIU T R, CHEN M. Analysis of foreign gene copy number in transgenic wheat by optimized digital PCR. Scientia Agricultura Sinica, 2020,53(10):1931-1939. (in Chinese)
[31]	KAUTBALLY S, LEPROPRE S, LERIGOLEUR A, GINION A, BEAULOYE C. Platelet acetyl-coa carboxylase phosphorylation: a risk stratification marker that reveals platelet-lipid interplay in coronary artery disease patients. Archives of Cardiovascular Diseases Supplements, 2019,11(2):185-186.
[32]	LIDA M, YAMASHIRO S, YAMAKAWA H, HAYAKAWA K, KURIBARA H, KODAMA T, FURUI S, AKIYAMA H, MAITANI T, HINO A. Development of taxon-specific sequences of common wheat for the detection of genetically modified wheat. Journal of Agricultural and Food Chemistry, 2005,53(16):6294-6300.
[33]	SINGH R, DUBEY A K, SANYAL I. Optimisation of adventitious shoot regeneration and agrobacterium-mediated transformation in Canna × Generalis (Canna Lily). Horticultural Plant Journal, 2019,5(1):39-46.
[34]	YIN Y C, HOU J M, TIAN S K, YANG L, ZHANG Z X, LI W D, LIU Y. Overexpressing chalcone synthase (CHS) gene enhanced flavonoids accumulation in Glycyrrhiza uralensis hairy roots. Botany Letters, 2020,167(2):219-231.
[35]	YANG L T, DING J Y, ZHANG C M, JIA J W, WENG H B, LIN W X, ZHANG D B. Estimating the copy number of transgenes in transformed rice by real-time quantitative PCR. Plant Cell Reports, 2005,23(10/11):759-763.
[36]	WHALE A S, HUGGETT J F, COWEN S, SPEIRS V, SHAW J, ELLISON S, FOY C A, SCOTT D J. Comparison of microfluidic digital PCR and conventional quantitative PCR for measuring copy number variation. Nucleic Acids Research, 2012,40(11):e82.
[37]	姜羽, 胡佳莹, 杨立桃. 利用微滴数字PCR分析转基因生物外源基因拷贝数. 农业生物技术学报, 2014,22(10):1298-1305.
	JIANG Y, HU J Y, YANG L T. Estimating the exogenous genes copy number of genetically modified organisms by droplet digital PCR. Journal of Agricultural Biotechnology, 2014,22(10):1298-1305. (in Chinese)
[38]	GAO F G, PFEIFER E, FARAH H, KARAMPINI E, DUA D, KAMAI N, CANE P, TOBAL K, SETHI T, SPICER J, MCCAUGHAN F. Microdroplet digital PCR: Detection and quantitation of biomarkers in archived tissue and serial plasma samples in patients with lung cancer. Journal of Thoracic Oncology, 2015,10(1):212-217.
[39]	HINDSON C M, CHEVILLET J R, BRIGGS H A, GALLICHOTTE E N, RUF I K, HINDSON B J, VESSELLA R L, TEWARI M. Absolute quantification by droplet digital PCR versus analog real-time PCR. Nature Methods, 2013,10(10):1003-1005.
[40]	HARBERD N P, BARTELS D, THOMPSON R D. Analysis of the gliadin multigene loci in bread wheat using nullisomic-tetrasomic lines. Molecular and General Genetics, 1985,198(2):234-242.
[41]	CASSIDY B G, DVORAK J, ANDERSON O D. The wheat low molecular weight glutenin genes: characterization of six new genes and progress in understanding gene family structure. Theoretical and Applied Genetics, 1998,96(6/7):743-750.
[42]	KORBEL J O, KIM P M, CHEN X Y, URBAN A E, WEISSMAN S, SNYDER M, GERSTEIN M B. The current excitement about copy-number variation: how it relates to gene duplications and protein families. Current Opinion in Structural Biology, 2008,18(3):366-374.
[43]	CHEN C, WANG W, YUAN J X, CHEN J, MOU L M. Analysis of HMW-GS and LMW-GS in spring wheat varieties and key parental materials cultivated in Gansu dryland. Acta Agriculturae Boreali- occidentalis Sinica, 2018,27(11):1598-1605.
[44]	LI Y Y, XIAO J H, WU J J, DUAN J L, LIU Y, YE X G, ZHANG X, GUO X P, GU Y Q, ZHANG L C, JIA J Z, KONG X Y. A tandem segmental duplication (TSD) in green revolution gene Rht-D1b region underlies plant height variation. The New Phytologist, 2012,196(1):282-291.
[45]	SUTTON T, BAUMANN U, HAYES J E, COLLINS N C, SHI B J, SCHNURBUSCH T, HAY A, MAYO G M, PALLOTTA M A, TESTER M A. Adelaide research and scholarship: boron toxicity tolerance in barley arising from efflux transporter amplification. American Association for the Advancement of Science, 2007,318(5855):1446-1449.
[46]	FRANCIA E, MORCIA C, PASQUARIELLO M, MAZZAMURRO V, MILC J A, RIZZA F, TERZI V, PECCHIONI N. Copy number variation at the HvCBF4-HvCBF2 genomic segment is a major component of frost resistance in barley. Plant Molecular Biology, 2016,92(1/2):161-175.
[47]	KNOX A K, DHILLON T, CHENG H M, TONDELLI A, PECCHIONI N, STOCKINGER E J. CBF gene copy number variation at Frost Resistance-2 is associated with levels of freezing tolerance in temperate-climate cereals. Theoretical and Applied Genetics, 2010,121(1):21-35.
[48]	PEARCE S, ZHU J, BOLDIZSÁR Á, VÁGÚJFALVI A, BURKE A, KIMBERLEY G C, GÁBOR G, DUBCOVSKY J. Large deletions in the CBF gene cluster at the Fr-B2 locus are associated with reduced frost tolerance in wheat. Theoretical and Applied Genetics, 2013,126(11):2683-2697.
[49]	AURORA D, MELULEKI Z, ADRIAN S T, PETER L, DAVID A L. Copy number variation affecting the Photoperiod-B1 and Vernalization- A1 genes is associated with altered flowering time in wheat (Triticum aestivum). PloS ONE, 2012,7(3):e33234.
[50]	ARMOUR J A, SISMANI C, PATSALIS P C, CROSS G. Measurement of locus copy number by hybridisation with amplifiable probes. Nucleic Acids Research, 2000,28(2):605-609.
[51]	NITCHER R, DISTELFELD A, TAN C T, YAN L L, DUBCOVSKY J. Increased copy number at the HvFT1 locus is associated with accelerated flowering time in barley. Molecular Genetics and Genomics, 2013,288(5):261-275.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

引物/探针名称 Primer/probe name	序列 Sequence (5′-3′)	退火温度 Annealing temperature (℃)
Glu-A3F	AGCAATCCCGCCATGAGTCA	60
Glu-A3R	AGTGGTGGTTTCGTACAACG
Glu-A3T	AATGTGTCTCCCAACCCCAACAGCAG
Glu-B3F	CAAGTCATCTTTAGCAAGCATCAGG	59
Glu-B3R	AAGGTCTTCATGGTGGACTAGTGTT
Glu-B3T	ATAGTAGCCAGGGCACCACCTCTTT
Glu-D3F	ACTAATCGAGCATATCCTAA	51
Glu-D3R	GTTACATTGGGTTAGGTTT
Glu-D3T	AGCCTATACAAGGTTCCAAACTCGG
Acc1F	GCAATGTAGCTGCGCTTCAC
Acc1R	CCTGCAACCGTGGATTAAGT
T-Acc1	ACAGTAGCAGCACCAACATAACCCACAGC

DNA浓度 DNA concentration (ng·μL^-1)	Ct值 Ct value			平均值 Average value	标准差 Standard deviation	方差 Variance	变异系数 Coefficient of variation (%)
DNA浓度 DNA concentration (ng·μL^-1)	1	2	3	平均值 Average value	标准差 Standard deviation	方差 Variance	变异系数 Coefficient of variation (%)
300	27.16	27.55	27.22	27.31	0.2100	0.0441	0.77
200	27.96	28.08	28.10	28.05	0.0757	0.0057	0.27
100	30.20	30.21	30.17	30.19	0.0208	0.0004	0.07
50	30.84	30.96	30.91	30.90	0.0603	0.0036	0.20
30	31.02	31.09	30.81	30.97	0.1457	0.0212	0.47

品种名 Sample	重复1 Repeat 1			重复2 Repeat 2			重复3 Repeat 3			变异系数 Coefficient of variation	拷贝数 Copy number
品种名 Sample	Ct (Glu-A3)	Ct (Acc1)	3X₀/R₀	Ct (Glu-A3)	Ct (Acc1)	3X₀/R₀	Ct (Glu-A3)	Ct (Acc1)	3X₀/R₀	变异系数 Coefficient of variation	拷贝数 Copy number
烟农15 Yangnong 15	27.95	28.55	2.52	28.21	28.97	2.86	27.89	28.63	2.82	6.80	3
安农92484W Annong 92484W	26.58	27.87	4.50	26.71	27.93	4.24	26.63	27.91	4.45	3.14	5
百农64 Bainong 64	27.45	28.33	3.18	27.06	27.96	3.25	27.37	28.18	3.00	4.10	3
小偃6号 Xiaoyan 6	27.41	28.41	3.51	27.11	28.09	3.47	27.30	28.14	3.09	6.91	4
周麦16 Zhoumai 16	27.01	27.57	2.35	26.64	27.35	2.80	26.59	27.25	2.69	8.98	3
02P67	26.81	27.46	2.66	26.74	27.22	2.32	26.46	27.01	2.46	6.90	3
邯郸6172 Handan 6172	27.50	27.92	2.19	27.58	28.32	2.83	27.52	28.12	2.53	12.72	3
周麦18 Zhoumai 18	27.84	28.60	2.87	27.62	28.36	2.83	27.57	28.18	2.55	6.34	3

Analysis of Copy Number Variation of Glu-3 Locus in Common Wheat

RichHTML

PDF

Abstract

Cite this article

share this article

Figures/Tables 10

References 51

Related Articles 15

Metrics

Comments

Recommended 0

[1]	CHEN JiHao, ZHOU JieGuang, QU XiangRu, WANG SuRong, TANG HuaPing, JIANG Yun, TANG LiWei, $\boxed{\hbox{LAN XiuJin}}$, WEI YuMing, ZHOU JingZhong, MA Jian. Mapping and Analysis of QTL for Embryo Size-Related Traits in Tetraploid Wheat [J]. Scientia Agricultura Sinica, 2023, 56(2): 203-216.
[2]	YAN YanGe, ZHANG ShuiQin, LI YanTing, ZHAO BingQiang, YUAN Liang. Effects of Dextran Modified Urea on Winter Wheat Yield and Fate of Nitrogen Fertilizer [J]. Scientia Agricultura Sinica, 2023, 56(2): 287-299.
[3]	XU JiuKai, YUAN Liang, WEN YanChen, ZHANG ShuiQin, LI YanTing, LI HaiYan, ZHAO BingQiang. Nitrogen Fertilizer Replacement Value of Livestock Manure in the Winter Wheat Growing Season [J]. Scientia Agricultura Sinica, 2023, 56(2): 300-313.
[4]	ZHAO HaiXia,XIAO Xin,DONG QiXin,WU HuaLa,LI ChengLei,WU Qi. Optimization of Callus Genetic Transformation System and Its Application in FtCHS1 Overexpression in Tartary Buckwheat [J]. Scientia Agricultura Sinica, 2022, 55(9): 1723-1734.
[5]	WANG HaoLin,MA Yue,LI YongHua,LI Chao,ZHAO MingQin,YUAN AiJing,QIU WeiHong,HE Gang,SHI Mei,WANG ZhaoHui. Optimal Management of Phosphorus Fertilization Based on the Yield and Grain Manganese Concentration of Wheat [J]. Scientia Agricultura Sinica, 2022, 55(9): 1800-1810.
[6]	TANG HuaPing,CHEN HuangXin,LI Cong,GOU LuLu,TAN Cui,MU Yang,TANG LiWei,LAN XiuJin,WEI YuMing,MA Jian. Unconditional and Conditional QTL Analysis of Wheat Spike Length in Common Wheat Based on 55K SNP Array [J]. Scientia Agricultura Sinica, 2022, 55(8): 1492-1502.
[7]	MA XiaoYan,YANG Yu,HUANG DongLin,WANG ZhaoHui,GAO YaJun,LI YongGang,LÜ Hui. Annual Nutrients Balance and Economic Return Analysis of Wheat with Fertilizers Reduction and Different Rotations [J]. Scientia Agricultura Sinica, 2022, 55(8): 1589-1603.
[8]	LIU Shuo,ZHANG Hui,GAO ZhiYuan,XU JiLi,TIAN Hui. Genetic Variations of Potassium Harvest Index in 437 Wheat Varieties [J]. Scientia Agricultura Sinica, 2022, 55(7): 1284-1300.
[9]	WANG YangYang,LIU WanDai,HE Li,REN DeChao,DUAN JianZhao,HU Xin,GUO TianCai,WANG YongHua,FENG Wei. Evaluation of Low Temperature Freezing Injury in Winter Wheat and Difference Analysis of Water Effect Based on Multivariate Statistical Analysis [J]. Scientia Agricultura Sinica, 2022, 55(7): 1301-1318.
[10]	GOU ZhiWen,YIN Wen,CHAI Qiang,FAN ZhiLong,HU FaLong,ZHAO Cai,YU AiZhong,FAN Hong. Analysis of Sustainability of Multiple Cropping Green Manure in Wheat-Maize Intercropping After Wheat Harvested in Arid Irrigation Areas [J]. Scientia Agricultura Sinica, 2022, 55(7): 1319-1331.
[11]	ZHI Lei,ZHE Li,SUN NanNan,YANG Yang,Dauren Serikbay,JIA HanZhong,HU YinGang,CHEN Liang. Genome-Wide Association Analysis of Lead Tolerance in Wheat at Seedling Stage [J]. Scientia Agricultura Sinica, 2022, 55(6): 1064-1081.
[12]	QIN YuQing,CHENG HongBo,CHAI YuWei,MA JianTao,LI Rui,LI YaWei,CHANG Lei,CHAI ShouXi. Increasing Effects of Wheat Yield Under Mulching Cultivation in Northern of China: A Meta-Analysis [J]. Scientia Agricultura Sinica, 2022, 55(6): 1095-1109.
[13]	CAI WeiDi,ZHANG Yu,LIU HaiYan,ZHENG HengBiao,CHENG Tao,TIAN YongChao,ZHU Yan,CAO WeiXing,YAO Xia. Early Detection on Wheat Canopy Powdery Mildew with Hyperspectral Imaging [J]. Scientia Agricultura Sinica, 2022, 55(6): 1110-1126.
[14]	ZONG Cheng, WU JinXin, ZHU JiuGang, DONG ZhiHao, LI JunFeng, SHAO Tao, LIU QinHua. Effects of Additives on the Fermentation Quality of Agricultural By-Products and Wheat Straw Mixed Silage [J]. Scientia Agricultura Sinica, 2022, 55(5): 1037-1046.
[15]	MA HongXiang, WANG YongGang, GAO YuJiao, HE Yi, JIANG Peng, WU Lei, ZHANG Xu. Review and Prospect on the Breeding for the Resistance to Fusarium Head Blight in Wheat [J]. Scientia Agricultura Sinica, 2022, 55(5): 837-855.