Table of Content

12 October 2024, Volume 50 Issue 10

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Evaluation of editing efficiency of different CRISPR-Cas12f systems

HUANG Ling-Zhi, FU Xiao, QI Xian-Tao, LIU Chang-Lin, XIE Chuan-Xiao, WU Peng-Hao, REN Jiao-Jiao, ZHU Jin-Jie

Acta Agronomica Sinica. 2024, 50(10):  2425-2434.  doi:10.3724/SP.J.1006.2024.43012

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CRISPR/Cas12f proteins belonging to the Type V-F family are reported to be only 1/4 to 1/3 the size of Cas9 protein molecules, providing a significant advantage in viral vector delivery. However, the CRISPR/Cas12f system for gene editing in plants has been reported to have lower editing activity, limiting its broader application in plant research. In this study, we compared the editing activities of OsCas12f, SpCas12f, and UnCas12f in three different systems: in vitro digestion, yeast, and transient expression in maize protoplasts. The results showed that the editing activities of OsCas12f and SpCas12f proteins were comparable in terms of in vitro digestion of Cas12f/sgRNA complexes, while no substrate digestion activity was detected for UnCas12f. In the yeast mutant eGFP expression restoration assay, OsCas12f exhibited an editing efficiency of over 95% at the two tested loci, which was comparable to Cas12i.3. On the other hand, SpCas12f achieved editing efficiencies of 1.63% and 3.20% at the two sites, respectively, representing the next highest effect. However, UnCas12f showed minimal editing activity. Furthermore, by transiently expressing maize protoplasts, we compared the editing efficiencies of OsCas12f and SpCas12f at endogenous maize loci. It was found that OsCas12f successfully mediated targeted editing at two loci with editing efficiencies of 2.72% and 1.97%, respectively, while SpCas12f only mediated targeted editing at one locus with an editing efficiency of 1.09%. Deletion of bases was the predominant type of mutation introduced by Cas12f proteins at the target loci, with deletion lengths ranging from -9 to -17 base pairs. These comprehensive results indicate that OsCas12f can serve as a versatile tool for developing plant microgene editors and related technologies.

Development of low-glutelin rice germplasm by gene editing technology

ZHOU Tian-Tian, TANG Zhao-Cheng, LI Xiao, ZHU Peng, DENG Jing-Jing, YANG Yu-Wen, ZHANG Bao-Long, GUO Dong-Shu

Acta Agronomica Sinica. 2024, 50(10):  2435-2446.  doi:10.3724/SP.J.1006.2024.32060

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Rice (Oryza sativa L.) is a crucial cereal crop worldwide, with protein being its second-most significant nutritional component. Patients with kidney diseases are required to limit their protein intake to alleviate the metabolic burden on their kidneys and control disease progression. In regular rice cultivars, glutelin is the predominant protein component and is easily digested by the human body. In this study, simultaneous mutations were introduced into Glutelin A1 (GluA1), GluA2, and GluA3 using CRISPR/Cas9-mediated targeted mutagenesis in a japonica rice cultivar derived from Low Glutelin Content-1 (LGC-1). Consequently, a low-glutelin rice germplasm with approximately 1.8% glutelin content, free from transgenic elements, was generated. The quality and agronomic traits of this germplasm were further comprehensively evaluated. The low-glutelin germplasms generated in this study exhibited significantly lower chalkiness degree compared to the recipient cultivar, while the brown rice rate and milled rice rate were significantly higher. This study presents a highly efficient and convenient method for generating rice germplasm with reduced glutelin content and offers new genetic materials for the cultivation of functional rice cultivars suitable for patients with kidney diseases.

Transcription factor GhWRKY41 enhances cotton resistance to Verticillium dahliae by promoting SA synthesis

XIAO Sheng-Hua, DONG Xian-Man, PENG Xin, LI An-Zi, BI Zhao-Fu, LIAO Ming-Jing, HUANG Li-Hao, GUAN Qian-Qian, HU Qin, ZHU Long-Fu

Acta Agronomica Sinica. 2024, 50(10):  2447-2457.  doi:10.3724/SP.J.1006.2024.44009

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Cotton is an important commercial crop, and its yield and quality are severely affected by Verticillium dahliae. Identifying cotton resistance genes to Verticillium dahliae and exploring the underlying molecular mechanisms is of great significance for accelerating the breeding process of cotton resistant to Verticillium wilt. In a previous study, the WRKY gene GhWRKY41 was identified as being induced by Verticillium dahliae in multiple resistant cotton varieties, enhancing cotton resistance through the activation of phenylpropanoid metabolism. This study further analyzed the expression patterns of GhWRKY41 under different hormone treatments, validated its disease resistance function in the upland cotton variety ‘Jin668’ using a virus-induced gene silencing (VIGS) assay, and measured endogenous hormone content. The results demonstrated that GhWRKY41 was significantly up-regulated by SA, Me-JA, and H₂O₂. Silencing GhWRKY41 weakened cotton resistance to Verticillium dahliae, while overexpression of GhWRKY41 led to a marked increase in SA content, and its RNAi lines showed a decrease in SA content. RT-qPCR results revealed that the expression levels of the SA biosynthesis gene GhSID2 and the SA signal transduction genes GhNPR1, GhPR1, and GhPR5 were significantly up-regulated in GhWRKY41-overexpression plants but decreased in GhWRKY41-RNAi plants. ChIP-qPCR and luciferase reporter gene assays showed that GhWRKY41 binds to and activates the expression of GhSID2, GhPR1, and GhPR5. Additionally, external spraying of SA significantly enhanced cotton resistance to Verticillium dahliae. In summary, GhWRKY41 enhances cotton resistance to Verticillium dahliae by promoting SA synthesis. This study elucidates the biological function of GhWRKY41 in cotton resistance to Verticillium dahliae and provides a theoretical basis for developing cotton varieties with improved resistance.

Overexpression of wild soybean salt-alkali tolerance gene GsGSTU13 increases salt-alkaline tolerance in rice seedlings

LI Wan-Hong, HU Bing-Shuang, SUN Xiao-Li, CAI Xiao-Xi, SUN Ming-Zhe

Acta Agronomica Sinica. 2024, 50(10):  2458-2467.  doi:10.3724/SP.J.1006.2024.44017

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Glutathione S-transferases (GSTs) are a class of highly conserved enzymes that play crucial roles in plant responses to environmental stresses. Bioinformatic analysis has revealed that Glycine soja GsGSTU13, which positively regulates salt-alkaline tolerance, shares the highest sequence identity with the OsGSTU17 protein. To investigate the potential contribution of GsGSTU13 to rice salt-alkaline tolerance, we transformed GsGSTU13 into rice and obtained two homozygous transgenic lines with significantly elevated GST activity. Phenotypic assays showed that after treatment with 200 mmol L^-1 NaHCO₃, the accumulation of reactive oxygen species was significantly lower in GsGSTU13 transgenic lines compared to wild-type. Additionally, the survival rates, relative water contents, and the activities of superoxide dismutases, peroxidases, catalases, and GSTs were significantly higher in GsGSTU13 transgenic lines than in the wild-type. In summary, overexpression of GsGSTU13 in rice enhanced salt-alkaline tolerance by promoting ROS scavenging, which could facilitate the breeding of new rice cultivars with improved tolerance to salt-alkaline stress.

Analysis of agronomic, quality traits and genetic diversity of domestic and foreign foxtail millet resources

XUE Ya-Peng, XIN Xu-Xia, WANG Ruo-Nan, YU Xiao-Han, LIU Shao-Xiong, WANG Rui-Yun, LIU Min-Xuan

Acta Agronomica Sinica. 2024, 50(10):  2468-2482.  doi:10.3724/SP.J.1006.2024.44041

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The present study aimed to investigate the comprehensive agronomic and quality traits of newly introduced foreign foxtail millet resources and their compatibility with domestic local varieties. The data obtained will serve as a reference for the identification and utilization of outstanding resources. In this research, a total of 88 newly introduced foxtail millet varieties and 152 domestic local varieties were selected for evaluation. Field assessments were conducted to examine agronomic traits, while quality analysis of harvested seeds was performed alongside kinship analysis using SNP markers derived from resequencing. The findings revealed significant differences in quantitative traits among the evaluated materials, indicating a rich genetic diversity. Seven traits exhibited a genetic diversity index (H') of ≥ 2.00. The top three materials, based on the comprehensive evaluation of quantitative traits, were Banyueying from Shandong province (F = 2.47), newly introduced germplasm Y-190 from the United Kingdom (F = 2.45), and Dabaigu I from Shanxi province (F = 2.38). In contrast, the differences in quality traits were not significant, with a genetic diversity index (H') ranging from 0.38 to 1.37. Correlation analysis demonstrated positive associations between most quantitative traits, except for spike thickness and thousand grain weight, which exhibited a non-significant negative correlation. The quality measurement results indicated that each trait followed a normal distribution and varied among different varieties. The genetic diversity index (H') of quality traits ranged from 1.65 to 2.05, with protein and fat exhibiting the highest genetic diversity (H' ≥ 2.00). Correlation analysis revealed a non-significant negative correlation between protein content and fat content, as well as between fat content and fiber content. Significant positive correlations were observed between starch content and fiber content, as well as L^* and b^* values. Based on phenotypic traits, the participating materials were classified into four groups, each showing distinct phenotypic characteristics. Furthermore, the SNP-based evolutionary tree and group structure analysis indicated various kinship relationships among different varieties. Notably, some newly introduced germplasms exhibited closer kinship ties with domestic local varieties. However, further research is required to elucidate the specific kinship relationships among different materials. This comprehensive evaluation of recently introduced foreign foxtail millet resources encompassed agronomic traits, quality traits, and their similarities and differences with domestic resources. The findings provide a foundation for the subsequent utilization and promotion of exceptional resources.

Genome-wide association analysis and candidate genes prediction of β-glucan content in barley grains

LU Zong-Hui, SI Er-Jing, YE Pei-Yin, WANG Jun-Cheng, YAO Li-Rong, MA Xiao-Le, LI Bao-Chun, WANG Hua-Jun, SHANG Xun-Wu, MENG Ya-Xiong

Acta Agronomica Sinica. 2024, 50(10):  2483-2492.  doi:10.3724/SP.J.1006.2024.31084

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A total of 238 barley germplasm resources from various sources were genotyped using a high-throughput SNP chip. The β-glucan content of barley grain was determined for two consecutive years. Genome-wide association analysis was conducted using a general linear model based on the PCA model. The results showed that the β-glucan content of the 238 barley materials ranged from 1.23% to 6.55% and 1.79% to 6.64% for the respective years, exhibiting a normal distribution. In the GWAS analysis, a total of 19 significant SNP markers were detected, located on chromosomes 1H, 2H, 3H, 4H, and 5H. These markers accounted for 7.39% to 10.29% of the observed phenotypic variation. Candidate genes were identified within a 300 kb range upstream and downstream of the SNP sites showing significant associations, and a total of 37 genes were identified. Through a combination of previous studies and BLAST gene annotation, four candidate genes most likely involved in β-glucan synthesis were identified. Notably, within 89 kb upstream of the most significant SNP locus B1_1033963, the candidate gene HORVU.MOREX.R3.1 HG0000140 was detected, which may play a crucial role in the process of beta-glucan synthesis. This study provides theoretical guidance and valuable gene resources for the genetic improvement of β-glucan content in barley.

QTL mapping for salt tolerance traits throughout the entire growth period of deep-water rice

XIA Xiu-Zhong, ZHANG Zong-Qiong, NONG Bao-Xuan, FENG Rui, GUO Hui, CHEN Can, LIANG Shu-Hui, ZHUANG Jie, LIAO Zu-Yu, SONG Guo-Xian, YANG Xing-Hai, LI Dan-Ting

Acta Agronomica Sinica. 2024, 50(10):  2493-2502.  doi:10.3724/SP.J.1006.2024.42007

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Salinity stress is a major constraint to rice production in many coastal regions, particularly in the salt-fresh water interface areas. Salt tolerance in rice is a complex trait that can be localized through quantitative trait loci (QTL) mapping, which can facilitate the breeding of rice varieties with enhanced salt tolerance. In this study, an F₉ mapping population consisting of 174 recombinant inbred lines was developed from a cross between the coastal deep-water rice variety Chihe (donor parent) and the U.S. rice variety Lemont (receptor parent). The population was subjected to salinity stress during the germination, seedling, and reproductive stages with NaCl concentrations of 15 g L^-1, 5 g L^-1, and 5-6 g L^-1, respectively. Data were collected on relative germination rates, seedling salt tolerance grades, and seven phenotypic traits during the reproductive stages. Linkage genetic mapping and QTL mapping were performed using 142 simple sequence repeat (SSR) markers. The results indicated that Chihe was salt-sensitive at the germination stage but exhibited salt tolerance at the seedling and reproductive stages, while Lemont lines were consistently salt-sensitive. Approximately 70.11%, 50.57%, and 60.34% of the lines showed salt sensitivity in the germination, seedling, and reproductive stages, respectively, with a weak negative correlation of salt tolerance across the stages. A total of 33 QTLs were identified, with LOD values ranging from 2.52 to 10.32 and phenotypic variation explained ranging from 0.06% to 13.68%. Specifically, 4 QTLs were identified for the germination stage, 6 for the seedling stage, and 23 for the reproductive stage. Four overlapping QTLs were identified at the reproductive stage, and the QTLs contributing to the largest phenotypic variation were all derived from the salt-tolerant parent. Further investigation of these QTLs can provide new genetic resources for improving salt tolerance in rice breeding, thereby aiding the development of rice varieties with enhanced tolerance to salinity stress.

Salt tolerance analysis of HvMBF1c in barley

ZHAN Xiao-Xiao, FENG Ju-Ling, ZHANG Zhen-Huan, ZHANG Hong, WANG Jun-Cheng, LI Bao-Chun, YANG Ke, SI Er-Jing, MENG Ya-Xiong, MA Xiao-Le, WANG Hua-Jun, YAO Li-Rong

Acta Agronomica Sinica. 2024, 50(10):  2503-2514.  doi:10.3724/SP.J.1006.2024.31075

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The HvMBF1c gene in barley is implicated in the plant’s response to salt stress. To investigate the HvMBF1c response mechanism to salt stress, we conducted bioinformatics analysis of the HvMBF1 gene, qRT-PCR analysis, and physiological index determination in Nakagawa barley (salt-tolerant type), GN18 (salt-sensitive type), wild-type (WT) and transgenic HvMBF1c Arabidopsis thaliana treated with 200 mmol L^-1 NaCl for 0, 6, 12, 24, 48, and 72 hours. The results showed that the barley HvMBF1c gene is closely related to the wheat TaMBF1c gene, and is located on chromosome 7H. The exon of HvMBF1c is larger than those of HvMBF1a and HvMBF1b, with HvMBF1a and HvMBF1b containing a typical motif 4 domain, while the HvMBF1c gene has a unique motif 5 domain. The promoter sequence of the HvMBF1c gene includes photoreactive, root-specific regulatory elements, and hypoxia-specific inducible enhancing elements, homologous to wheat chromosomes 2, 3, and 7. qRT-PCR analysis revealed significant upregulation of HvMBF1c expression in two barley genotypes at the seedling, jointing, heading, and filling stages, with higher expression in Nakagawa barley compared to GN18 as salt treatment duration increased. Similarly, the expression of HvMBF1c in Arabidopsis thaliana also increased with prolonged salt treatment time. Physiological indices of Nakagawa barley at each growth stage were lower than those in GN18, and transgenic Arabidopsis exhibited lower indices compared to WT under different salt stress conditions. These findings provide a foundation for further exploration of the HvMBF1c gene function.

Divergent evolutionary pattern of P5CS gene family and drought tolerance verification of LusP5CS1 in linseed

WANG Ling, ZHANG Yan-Ping, QI Yan-Ni, WANG Lei, LI Yu-Xiao, TAN Mei-Lian, WANG Wei

Acta Agronomica Sinica. 2024, 50(10):  2515-2527.  doi:10.3724/SP.J.1006.2024.34149

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Pyrroline-5-carboxylate synthetase (P5CS), a crucial enzyme encoded by the P5CS gene, plays a key role in the drought stress response in plants, primarily by regulating proline biosynthesis. In this study, we used two P5CS genes from Arabidopsis thaliana as query sequences to meticulously screen and identify members of the P5CS gene family from the genomes of various important grain and oil crops, including linseed, rapeseed, soybean, peanut, sunflower, rice, and wheat. By thoroughly analyzing the selection pressure, specific sites, and functional differentiation potential of P5CS genes across these diverse crops, we gained a comprehensive understanding of their evolutionary patterns. Furthermore, we validated the functions of these genes in Arabidopsis thaliana, a model organism. Our findings revealed significant differences in gene structure and evolutionary patterns among members of the linseed P5CS gene family compared to their homologs in other crops. Notably, overexpressing the positively selected linseed LusP5CS1 gene in Arabidopsis thaliana resulted in a substantial increase in proline accumulation and enhanced drought resistance in the transgenic plants. Interestingly, even under non-drought stress conditions, the transgenic Arabidopsis thaliana exhibited a notable fitness advantage. This groundbreaking study not only enhances our understanding of the molecular mechanisms underlying drought resistance but also provides a solid theoretical foundation for breeding drought-tolerant linseed varieties.

Genetic diversity analysis and evaluation of domestic and international cotton germplasm resources under different planting environments

SHANG Hong-Yan, PU Jing, KE Hui-Feng, GU Qi-Shen, SUN Zheng-Wen, YANG Jun, WANG Guo-Ning, ZHANG Yan, LU Huai-Yu, XU Dong-Yong, WU Li-Qiang, MA Zhi-Ying, WANG Xing-Fen, WU Jin-Hua

Acta Agronomica Sinica. 2024, 50(10):  2528-2537.  doi:10.3724/SP.J.1006.2024.44012

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Currently, there are several issues in cotton breeding in China, including the high uniformity of existing varieties, declining genetic diversity of germplasm resources, and underutilization of valuable gene resources. In light of these challenges, a comprehensive assessment and analysis of genetic diversity were conducted on 415 cotton germplasm resources collected from three major cotton regions in China (Yellow River Basin, Yangtze River Basin, and Northwest Inland) as well as international sources. The evaluation encompassed three yield traits and seven fiber quality traits in three different locations: Sanya city, Hainan province; Hejian city, Hebei province; and Xinji city, Hebei province. The findings revealed that Hainan exhibited the highest fiber weight per boll and lint percentage but had relatively poor fiber quality. Hejian displayed the highest boll weight and superior fiber quality, while Xinji had the lowest yield performance. Additionally, it was observed that 10 phenotypic traits demonstrated significant variation and abundant genetic diversity across all three environments. Notably, there were significant differences among cotton germplasm resources from different origins, except for fiber length, uniformity, and short fiber percentage. Germplasm resources from the Yellow River Basin showed the best yield traits, highest fiber length, and fiber strength, with the largest proportion of materials surpassing the ‘double 30’ threshold. However, these resources had relatively high Micronaire values. Germplasm resources from the Yangtze River Basin exhibited higher fiber weight per boll and lint percentage. Foreign germplasm resources displayed higher boll weight but the lowest lint percentage. Correlation and cluster analyses demonstrated predominantly positive associations among yield traits, leading to the classification of germplasm resources into five categories. Furthermore, a factor analysis was conducted to comprehensively rank the tested materials, resulting in the identification of several elite resources. These included seven germplasm resources with large boll weight ( > 7 g), 26 with high lint percentage ( > 42%), 11 with high fiber length and strength surpassing the ‘30’ threshold, and nine resources exhibiting excellent comprehensive traits. These findings provide valuable parental materials for cotton breeding and serve as a crucial foundation for further research endeavors.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY

Photosynthetic repair of Glycine max (Linn.) Merr. by compound fungus agents and immobilization effect under cadmium stress

LIU Hong-Yuan, CEN Kai, LIU Yi-Lin, LOU Xue-Yi, ZHANG Ya-Ting, WU Jia-Rui, TAN Yu-Yu, ZHU Jia-Cheng, FANG Fang, LIU Peng

Acta Agronomica Sinica. 2024, 50(10):  2538-2549.  doi:10.3724/SP.J.1006.2024.44046

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Cadmium, a highly toxic heavy metal affecting food crops like soybean, not only inhibits plant growth and development but also damages the photosynthetic system, leading to reduced photosynthesis rates. Current cadmium remediation technologies primarily focus on the application of plant hormones and alteration of planting patterns, while the interaction between microorganisms and plants remains underexplored. This study aims to explore the remediation potential of compound white rot fungi in addressing cadmium pollution and the practical application value of immobilization technology. Four types of white rot fungi and Glycine max (Linn.) Merr. were used to prepare solid bacterial agents and set up a soil cultivation method for soybeans. We simulated cadmium-contaminated soil concentrations of 0, 50, and 100 mg L^-1. Three treatments were conducted for each concentration: a control group (CK) with no treatment, an experimental group with free strains (EG1), and an experimental group with solid agents (EG2). We examined the effects of mixed fermentation and immobilization technology on the strains’ adsorption efficiency and established the correlation between cadmium toxicity, immobilized microspheres, and soybean plants. The results indicate that, except for Phanerochaete chrysosporium, the other three strains demonstrated good compatibility. A mixed group of bacterial strains containing Pleurotus sajor-caju and Coriolus versicolor in a 1:1 ratio achieved an adsorption rate of 87.33% in cadmium-contaminated solutions at a concentration of 50 mg L^-1. To prolong the duration and improve the adsorption effect of the mixed strain, PVA mixed pellets with a sodium alginate (SA) concentration of 10 g L^-1, biochar mass concentration of 15 g L^-1, and bacterial content of 2% achieved a degradation rate of (95.12 ± 1.68)% within 96 hours after adding appropriate additives. Introducing immobilized mixed bacteria into simulated cadmium-contaminated soil inhibited the growth and photosynthetic indices of soybeans. The maximum decrease in F_o was 42.5%, and the maximum increase in F_v/F_m was 17.2%. After 14 days, the soybean antioxidant system was enhanced, with the highest activities of SOD, POD, and CAT being 27.34%, 12.41%, and 13.58%, respectively, in the CK group. Additionally, there was an increase in proline content and a decrease in malondialdehyde content, indicating enhanced plant resistance. In conclusion, cadmium stress suppresses the photochemical reaction center II in plants’ photosystems. The immobilization of mixed strains results in higher adsorption efficiency compared to single or free states. Applying a reliable bacterial agent enables soybeans to effectively trigger their light protection mechanism, produce osmoregulatory substances, and activate the antioxidant system, thus maintaining a stable redox environment and coping with cadmium stress.

Exploring the functions of different leaf types of directly-sown rapeseed under high density

HUANG Xiao-Yu, LOU Hong-Xiang, SHAO Dong-Li, ZHANG Zhe, JIANG Bo, XIAO Ya-Dan, CHANG Ying, GUO An-Da, ZHAO Jie, XU Zheng-Hua, WANG Jing, WANG Bo, KUAI Jie, ZHOU Guang-Sheng

Acta Agronomica Sinica. 2024, 50(10):  2550-2561.  doi:10.3724/SP.J.1006.2024.44032

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Optimal planting density is a core technology for ensuring rapeseed yield and production efficiency. However, excessive planting density can worsen lodging of rapeseed and make it challenging to achieve high yields. Leaves play a vital role in photosynthesis and dry matter accumulation in rapeseed, but the effects of different leaf types on yield and lodging resistance under high-density conditions remains unclear. In this study, we selected Zhongshuang 11 and implemented two planting densities (D3: 4.5×10⁵ plants hm^-2, D5: 7.5×10⁵ plants hm^-2) and four-leaf pruning treatments (CK: no leaf pruning, LP: pruning half of the long-petiole leaves, SP: pruning half of the short-petiole leaves, SL: pruning half of the sessile leaves). We aimed to investigate the effects of these three leaf types on the yield and lodging resistance of directly-sown rapeseed under high density, providing a theoretical basis for managing the source-sink relationship, high-density rapeseed cultivation, and the lodging challenge. The results showed that, at the same density, the proportions of short-petiole leaf > long-petiole leaf > sessile leaf. When the density increased from D3 to D5, about 2 short-petiole leaves were reduced, resulted in a decrease in proportion from 50.6% to 46.7%. Leaf pruning at both densities lead to a reduction in siliques per plant, consequently decreasing the yield per plant and actual yield. Among the pruning treatments, SP had the greatest impact on yield. As density increased, the effect of SP on yield per plant increased from 19.61% to 37.67%, and the effect on actual yield increased from 13.65% to 22.03%. The lodging index increased under different leaf pruning treatments, with SP treatment significantly increasing the lodging index in both the upper and lower parts of the plant, indicating weaker lodging resistance. In addition, the root surface area and volume per plant decreased with increasing density, particularly under LP and SP treatments. Correlation analysis revealed that, under D3 density, pruning short-petiole leaves reduced the number of siliques per plant. Under D5 density, pruning short-petiole leaves and long-petiole leaves resulted in reduced siliques per plant and seed per silique, significantly decreasing both yield and lodging resistance. Pruning long-petiole leaves led to a significant decrease in root surface area and volume, which was unfavorable for yield formation.

Response of the yield sustainability of maize with different irrigated quota to intercropped green manure

WANG Yun-Jie, FAN Zhi-Long, ZHANG Diao-Liang, MAO Shou-Fa, HU Fa-Long, YIN Wen, CHAI Qiang

Acta Agronomica Sinica. 2024, 50(10):  2562-2574.  doi:10.3724/SP.J.1006.2024.33068

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Clarifying the effect of intercropped green manure on the yield sustainability of maize under different irrigation levels plays a crucial role in the development of a water-saving and efficiency-oriented maize and leguminous green manure intercropping system. Field experiments were conducted from 2018 to 2022 using a split-plot experimental design, including two planting patterns: maize and green manure intercropping (M/V) and sole maize cropping (SM). Three irrigation levels were implemented: local conventional irrigation (I₃: 4000 m³ hm^-2), 15% reduction in irrigation (I₂: 3400 m³ hm^-2), and 30% reduction in irrigation (I₁: 2800 m³ hm^-2). The effects of intercropped green manure on maize yield, yield components, relative multi-crop resistance index, yield sustainability index, key soil nutrient indicators, and economic benefits under different irrigation levels were investigated. The results indicated that intercropped green manure increased the number of maize kernels and thousand-kernel weight when subjected to a 15% reduction in irrigation, compared to sole maize cropping, resulting in enhanced grain yield. From 2019 to 2022, intercropped maize with a 15% reduction in irrigation exhibited an increase in the number of maize kernels and thousand-kernel weight by 8.7%-16.4% and 7.1%-13.4%, respectively, compared to sole maize cropping. Grain yield also increased by 9.3%-23.6%. There was no significant difference in grain yield between intercropped maize with a 15% reduction in irrigation and local conventional irrigation. The relative multi-crop resistance index of intercropped maize under reduced irrigation was greater than 0 and increased with the duration of planting, reducing the coefficient of variation in yield and increasing the yield sustainability index of maize. Under the same irrigation levels, the coefficient of variation in yield of intercropped maize decreased by 77.9%-82.8%, and the sustainability index increased by 12.2%-19.9% compared to sole maize cropping. Maize intercropped with green manure also led to increased soil nutrient levels compared to sole maize cropping. Soil organic matter and available nitrogen content increased by 6.4%-15.8% and 12.1%-35.6%, respectively. Soil available phosphorus content in maize and green manure intercropping with a 15% and 30% reduction in irrigation increased by 19.4% and 11.3%, respectively, compared to sole maize cropping. The content of soil organic matter, available nitrogen, and available phosphorus in maize and green manure intercropping with a 15% reduction in irrigation showed no significant difference compared to local conventional irrigation but was significantly greater than other treatments. The net income of intercropped maize with a 15% reduction in irrigation increased by 10.5%-34.2% compared to sole maize cropping from 2020 to 2022. From 2021 to 2022, the investment ratio of intercropped maize with a 15% reduction in irrigation increased by 7.8%-10.4% compared to sole maize cropping and showed no significant difference compared to intercropped maize with local conventional irrigation. Intercropped green manure improved the yield sustainability of maize by increasing soil organic matter and improving soil nitrogen and phosphorus conditions. In summary, intercropped green manure can mitigate the yield loss of maize caused by reduced irrigation, enhance the stability and sustainability index of maize under reduced irrigation. In this investigation, maize and common vetch intercropping with an irrigation level of 3400 m³ hm^-2 can serve as a reference for sustainable maize production and suitable irrigation levels in the experimental region.

Effect of light intensity on leaf photosynthetic physiology and root system of sweet potato in the early stage of growth

JIANG Yang-Ying, TANG Ming-Jun, ZHANG Lin-Xi, LYU Chang-Wen, TANG Dao-Bin, WANG Ji-Chun

Acta Agronomica Sinica. 2024, 50(10):  2575-2585.  doi:10.3724/SP.J.1006.2024.44011

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This study investigated the impact of different light intensities on leaf photosynthetic physiology and tuberization during the early growth stage of sweet potatoes. The findings contribute to theoretical and practical strategies for achieving high yields through relay intercropping of high-position crops with sweet potatoes. To explore the photosynthetic characteristics, tissue structure, and tuberization of sweet potato leaves, a two-factor split plot experiment was conducted in 2021. The main plot consisted of three sweet potato cultivars with varying root drying rates and leaf types: S1 (Chaoshu 1), S2 (Guangshu 87), S3 (Yusu 162). The subplot included three light intensities: L200 [(200 ± 50) μmol m^-2 s^-1], L500 [(500 ± 50) μmol m^-2 s^-1], L800 [(800 ± 50) μmol m^-2 s^-1]. The results revealed that decreasing light intensity led to reductions in upper epidermis thickness, palisade tissue thickness, spongy tissue thickness, leaf thickness, net photosynthetic rate (P_n), transpiration rate (T_r), stomatal conductance (G_s), RUBP carboxylase activity, number of storage roots per plant, storage root weight per plant, and dry matter weight of storage roots. Among these parameters, the treatment with high light intensity (L800) exhibited the best performance. However, the intercellular CO₂ concentration (C_i), chlorophyll a content, chlorophyll b content, carotenoid content, chlorophyll a/b ratio, and storage root drying rate were optimal under low light intensity (L200). Comprehensive evaluation through factor analysis revealed that the S3 variety with high root drying rate performed best under low light intensity, while the S1 variety with low root drying rate performed the worst. Sweet potato leaves primarily enhance light energy capture by increasing the content of photosynthetic pigments and leaf area index. They adapt to low light environments through plasticity in leaf anatomical structure, photosynthetic physiology, and RUBP carboxylase activity.

Effect of high-density planting and fertilizer reduction on yield and growth and development of indica hybrid rice

LU Jian-Xiang, GAO Qian-Wen, GAO Zhi-Qiang, YANG Hui-Bing, WEN Shuang-Ya, SHI Nan, HU Wen-Rui, JIN Yu-Hao, CHEN Long, LIU Yun, CAO Zheng-Deng-Yuan

Acta Agronomica Sinica. 2024, 50(10):  2586-2598.  doi:10.3724/SP.J.1006.2024.32051

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To investigate the effects of increased planting density and reduced fertilizer application on the biological traits, accumulation of dry matter, and yield of rice, this study employed three transplanting densities: low density (LD), medium density (MD), and high density (HD) with 18×10⁴, 22×10⁴, and 27×10⁴ plants hm^-2, respectively. Three fertilizer application levels were also set: low fertility (LF), medium fertility (MF), and high fertility (HF) with rates of 450, 525, and 675 kg hm^-2, respectively. Field experiments were conducted using sequential machine throwing technology during the period from 2021 to 2022. The results demonstrated that the medium density and medium fertilizer (MDMF) treatment exhibited the highest yield. Compared to the low density and high fertilizer (LDHF) treatment, the yield of MDMF treatment increased by 3.15% on average in two years. This treatment significantly increased the effective panicle number and stabilized other yield factors. However, there was no significant increase, and possibly a negative effect, on the harvest index. Prior to full heading, the shoot dry matter treated with high density and high fertilizer (HDHF) exhibited the highest quality, with an average increase of 0.17% over two years. On the other hand, the shoot dry matter treated with MDMF showed the highest quality after full heading, with an average increase of 0.16%. The leaf area index (LAI) and soil and plant analyzer development (SPAD) values of the MDMF treatment did not differ significantly from those of the LDHF treatment at each growth stage. However, the SPAD attenuation rate and LAI attenuation rate of the MDMF treatment remained consistently high. The SPAD attenuation rates were 12.65% and 16.85%, respectively, and the decay rates of LAI were 6.42% and 6.74%, respectively. The number of rice tillers increased with higher transplanting density and fertilizer application. In summary, appropriate densification and fertilizer reduction can increase the number of effective panicles, establish a more robust population structure and aboveground dry matter weight, enhance the production and transport capacity of indica hybrid rice from the full heading stage to the mature stage, maintain source stability, improve yield potential, and achieve high yields. The recommended combination for the production of indica hybrid rice is a planting density of 22×10⁴ plants hm^-2 and a fertilizer application rate of 525 kg hm^-2.

Effects of climate change and crop rotation system on forage oats yield in northern Shanxi province

CHENG Hua-Qiang, HOU Qing-Qing, ZHU Min, YANG Xuan

Acta Agronomica Sinica. 2024, 50(10):  2599-2613.  doi:10.3724/SP.J.1006.2024.31082

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To investigate the suitability of crop rotation systems involving forage oats (Avena sativa L.) and their response to climate change in northern Shanxi province, this study utilized the validated APSIM (Agricultural Production System sIMulator) climate model. Three planting patterns were employed: O-O (continuous cropping of forage oats), P-O (rotation of potato and forage oats), and M-O (rotation of maize and forage oats). Scenario simulations were conducted at eighteen sites. The results demonstrated the effective simulation of maize, potato, and forage oats production in northern Shanxi Province using APSIM. The normalized root mean square error (NRMSE) values were below 21%, while the Willmott agreement index (d) values exceeded 0.90. The highest forage oats yield was observed in Pinglu, Shenchi, and Zuoyun, ranging from 16,020 to 20,817 kg hm^-2. The forage yield of O-O, M-O, and P-O systems in the mid-period (MID) exhibited an increase of 5.49% to 23.20% compared to the base period (BAS). In the end period (END), the forage oats yield in Daixian, Datong, and 10 other locations improved by 0.27% to 9.15% compared to the MID period, while it decreased by 1.84% to 4.10% in Zuoyun, Shuozhou, Youyu, and Shenchi. Notably, the forage yield of O-O in Fanshi was 22.76% lower in the END period compared to the MID period. The P-O system exhibited superior soil water retention compared to other systems after the growing seasons, making it more effective in achieving high and stable forage oats production in most cases. For the Yanggao site, which had poor soil water storage capacity but high plant available water, the projected high precipitation conditions would compensate for the relatively high-water consumption in the O-O system. Overall, the findings of this study contribute to understanding the response mechanism of forage oats production to regional climate change in northern Shanxi Province and provide a theoretical basis for the scientific management of high and stable forage oats production.

Simulating effects of climate change on growth season and yield of double cropping rice in Jiangxi province based on DSSAT model

ZHANG Fang-Liang, LIU Wen-Ying, TIAN Jun, WANG Jian-Jun, LIU Dan, YANG Jun, LI Ying-Chun, ZHANG Yi-Zhi

Acta Agronomica Sinica. 2024, 50(10):  2614-2624.  doi:10.3724/SP.J.1006.2024.32057

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Jiangxi province is a key region for double cropping rice cultivation in China. Climate change has significantly impacted double cropping rice production in this area. This study validates the DSSAT model using daily meteorological data and crop data for double cropping rice in Jiangxi Province from 1981 to 2022. The validated DSSAT model is then used to analyze the spatial distribution and temporal variation trends of the growth season and yield of double cropping rice in Jiangxi province. Additionally, the t-test method is employed to identify differences in the effects of climate change on early rice and late rice in the province. The results are as follows: (1) The normalized root mean square error (NRMSE) between simulated and observed values for the sowing-to-flowering duration, sowing-to-maturity duration, and yield of early rice (late rice) in Jiangxi province are 1.87% (1.86%), 2.05% (2.36%), and 6.05% (7.30%), respectively. The D-index for these parameters are 0.97 (0.98), 0.96 (0.96), and 0.95 (0.94), respectively. (2) With fixed sowing dates and varieties, the growth seasons for early rice and late rice in Jiangxi province significantly shortened from 1981 to 2022, with an average decrease of 2.22 and 1.61 days per decade, respectively. The potential yields of early rice and late rice also significantly decreased over the same period, with an average reduction of 181.30 kg hm^-2 and 276.16 kg hm^-2 per decade, respectively. (3) The t-test results indicate that the climate trend for the growth season of early rice in Jiangxi Province is significantly lower than that of late rice. Conversely, the climate trend for the potential yield of early rice is significantly higher than that of late rice. The DSSAT model effectively simulates the growth and yield of double cropping rice in Jiangxi province. The findings highlight that climate change has more pronounced effects on the growth season of early rice and the potential yield of late rice in Jiangxi province. This study provides a scientific basis for crop model research, yield prediction, and climate change assessment for double cropping rice in Jiangxi province.

Effect of planting uniformity with machine on yield construction and quality formation of rice

TANG Cheng-Han, CHEN Hui-Zhe, YE Tian-Cheng, ZHANG Yu-Ping, XIANG Jing, ZHANG Yi-Kai, WANG Zhi-Gang, WANG Ya-Liang

Acta Agronomica Sinica. 2024, 50(10):  2625-2636.  doi:10.3724/SP.J.1006.2024.42004

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Planting uniformity plays a crucial role in determining the yield and quality of rice. This experiment aimed to elucidate the mechanism behind the simultaneous improvement of uniformity, yield, and quality. Three varieties were used in the study: Yongyou 1540 (indica-japonica hybrid rice), Zhehexiang 2 (inbred japonica rice), and Huazheyou 210 (hybrid indica rice). Four treatments with varying planting uniformity were established: T₁ (drill sowing, planting uniformity of 65%-75%), T₂ (broadcast sowing, planting uniformity of 45%-55%), T₃ (manual simulated mechanical rice transplanting, planting uniformity of 100%), and T₄ (manual simulated mechanical rice transplanting, planting uniformity of 50%). The study compared and analyzed the dynamic changes in tiller number, leaf area photosynthetic efficiency, dry matter accumulation and transport, yield formation, and rice quality across different planting uniformity groups. The results showed as follows: (1) Enhanced planting uniformity increased the number of productive tillers by promoting tillering. The average number of tillers at the tillering peak stage was highest in T₃, followed by T₁, T₂, and T₄, with a consistent trend across different varieties. (2) Enhanced planting uniformity led to an increase in population leaf area index, particularly in the highly effective leaf area index of the top three leaves at the heading stage. Additionally, improved uniformity enhanced dry matter accumulation and facilitated its transportation. Dry matter accumulation at maturity followed the order T₃ > T₁ > T₂ > T₄, with consistent trends observed among different varieties. However, there was no significant difference between T₂ and T₄. The effect of uniformity on stem sheath material movement during the filling stage varied among varieties, with no significant differences found between T₁, T₂, and T₄. (3) The number of grains per panicle did not significantly differ under different uniformity treatments. However, high planting uniformity resulted in a more uniform distribution of grains on primary and secondary branches. (4) Increased yield in high uniformity populations was primarily attributed to the effective panicle number, with consistent trends observed among different varieties. The average yield of T₃ was 8.16%, 15.41%, and 15.61% higher than that of T₁, T₂, and T₄, respectively. (5) Improving planting uniformity increased the brown rice rate, milled rice rate, head rice rate, and protein content, while decreasing chalkiness and chalky rice rate. These findings indicate that improving planting uniformity can promote tillering, increase leaf area index and dry matter accumulation, enhance effective panicle number, and ultimately improve yield and milled rice rate, as well as rice quality to some extent. The experimental results highlight the potential of precision sowing machines to improve rice yield and quality by enhancing planting uniformity.

RESEARCH NOTES

Molecular pyramiding of insect and glyphosate-resistant genes and correlation analysis on economic traits of the pyramided lines in upland cotton

CHEN Xu-Sheng, ZHAO Liang, DI Jia-Chun

Acta Agronomica Sinica. 2024, 50(10):  2637-2642.  doi:10.3724/SP.J.1006.2024.44028

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Breeding cotton with simultaneous resistance to cotton bollworm and glyphosate can enhance the ability of resistance to insects and herbicides, reduce pest control and herbicide costs, and improve comprehensive economic productivity in cotton production. In this study, a hybrid population was developed using self-bred high-yield and high-quality upland cotton, domestic transgenic Bt cotton lines, and GR79+GAT transgenic upland cotton lines as parental hybrids. Non-glyphosate-resistant cotton seedlings were eliminated by applying 0.2% glyphosate in the seedling beds, and cotton plants were screened for resistance to cotton bollworm under field conditions without pest control. Specific primers were used in the laboratory to track the insect-resistant and glyphosate-resistant genes, and individual plants carrying the Bt and GR79+GAT genes were propagated into plant lines. Nine lines exhibiting resistance to both insects and glyphosate were obtained through field screening. Subsequently, the economic traits of the selected lines were compared and analyzed. Among these lines, strain BG-6 demonstrated the combined presence of insect-resistant and glyphosate-resistant genes, as well as higher lint yield and fiber quality. The fiber length, specific strength, and micronaire value of BG-6 were 30.9 cm, 30.1 cN tex^-1, and 4.9, respectively. Correlation analysis between yield traits and fiber quality traits revealed a negative correlation between lint percentage and uniformity index (r = -0.838^**), but no significant correlation was observed between other yield traits and fiber quality traits. In conclusion, our study successfully selected resistant cotton seedlings through glyphosate spraying in the seedling beds, identified resistant cotton plants under field conditions without pest control, detected two resistance genes using PCR in the laboratory, and evaluated high-yield and high-quality traits in the field. By following this approach, we achieved the pyramiding of multiple traits, including insect resistance, glyphosate resistance, and high-yield and high-quality characteristics, in the segregated progeny of comprehensive hybrid combinations.

Molecular identification of broomcorn millet germplasm resources in Shaanxi based on SSR

GUO Juan, XIN Xu-Xia, FENG Zhi-Zun, CAO Yue, WANG Xiao-Dan, CAO Xiao-Ning, SANTRA Dipak K, CHEN Ling, QIAO Zhi-Jun, WANG Rui-Yun

Acta Agronomica Sinica. 2024, 50(10):  2643-2653.  doi:10.3724/SP.J.1006.2024.44037

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To construct the DNA molecular identity card of broomcorn millet (Panicum miliaceum L.), we conducted an experiment using 181 core germplasm samples of Shaanxi millets. The core primers were obtained through several rounds of PCR screening and optimization of millet-specific SSR markers developed in the preliminary stage of our research group. Chromosomal localization of the core markers was performed by comparing their sequences with the reference genome information of broomcorn millet using BLAST. FAM/HEX labels were added the 5' end of SSR primers, and the presence or absence of fragments detected by capillary electrophoresis was represented as “0/1”. The differentiation of fragments was carried out using ID Analysis 4.0, and the size of the amplified fragments was recorded in format (0-9) to construct unique material strings. These strings were then subjected to genetic diversity analysis using software tools such as Pop Gene, Power Marker, MEGA, Structure, and NTSYS. Our test results showed that seven combinations of fluorescent SSRs (RYW3, RYW6, RYW37, RYW40, RYW43, RYW125, and RYW146) could distinguish the 181 materials. These markers were unevenly distributed across five chromosomes, and a total of 77 allelic variants were detected, with an average of 11 variants per marker. The observed Shannon's diversity index (I) ranged from 0.8145 (RYW146) to 7.8254 (RYW125), with an average of 5.9076. The observed heterozygosity (Ho) ranged from 0.2627 (RYW146) to 0.9506 (RYW3), while the expected heterozygosity (He) ranged from 0.3329 (RYW146) to 0.8747 (RYW125). Nei’s gene diversity index (Nei) varied from 0.3315 (RYW146) to 0.8722 (RYW125). The Polymorphism Information Content (PIC) ranged from 0.5923 (RYW146) to 0.9445 (RYW125), with an average of 0.8419. Using the UPGMA clustering method, we categorized the 181 resources into three clusters. Principal component analysis resulted in the classification of the materials into ten taxa, which were consistent with their geographic sources. To provide convenient access to the DNA molecular identity cards, we utilized online QR code technology (https://cli.im/) for the 181 Shaanxi broomcorn millet core germplasm samples.

An E3 ubiquitin ligase gene TaSINA-3A is associated with plant height and 1000-grain weight in various environments in wheat

ZHAO Yang, LI Long, YANG Jin-Wen, JING Rui-Lian, SUN Dai-Zhen, WANG Jing-Yi

Acta Agronomica Sinica. 2024, 50(10):  2654-2664.  doi:10.3724/SP.J.1006.2024.41008

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The SINA (seven in absentia) family of E3 ubiquitin ligases is involved in plant growth and development, immune symbiosis and response to stresses. In this study, we cloned the TaSINA-3A gene located on chromosome 3A in wheat. The TaSINA-3A genome sequence spans 3897 bp, including a 2001 bp promoter region and a 1896 bp coding region. The TaSINA-3A genome encods 244 amino acids, and contains a conserved RING (really interesting new gene) domain at amino acids 50-91. Sequence polymorphism analysis revealed the presence of 33 single nucleotide polymorphisms (SNPs) in the promoter region and 4 SNPs in the coding region. Quantitative analysis showed that the TaSINA-3A gene exhibited tissue-specific expression during different developmental stages of wheat. Molecular markers based on the SNP loci SNP-159, SNP-418, and SNP-1286 in the promoter region were developed to genotype the natural population of wheat. Association analysis between genotypes and phenotypic traits revealed significant associations between molecular marker CAPS-159 and penultimate internode length (PIL) and plant height (PH) under various environmental conditions, and Association analysis between genotypes and phenotypic traits revealed significant associations between molecular marker dCAPS-418 and spike length (SL) and 1000-grain weight (TGW) under various environmental conditions. Among these associations, the allelic variant SNP-159-C was associated with shorter PIL and PH, while SNP-418-A was associated with longer SL and higher TGW. TaSINA-3A was found to negatively regulate spike and grain development, and its expression may be suppressed by the MYC transcription factor. In the context of wheat breeding in China, SNP-418-A has been positively selected and its frequency in modern cultivars has gradually increased; however, its full potential has not been fully utilized. These results provide valuable insights for further exploration of the mechanisms underlying penultimate internode length, plant height, spike length, and grain yield. Moreover, they provide genetic resources for breeding new varieties with high yield, stable yield and wide suitability.