Genetic transformation serves as a critical tool for gene function research and crop improvement. However, its efficiency is often low and highly dependent on species, genotypes, and explant types, significantly restricting its broader application. Many developmental factors have been proven pivotal not only for plant growth and development but also for the regulation of callus formation and shoot regeneration, which are key steps in the process of genetic transformation. Thus, this review focuses on the application of developmental factors in enhancing transformation efficiency across species. Developmental factors are classified into four regulatory pathways: morphogenesis, wound signaling, epigenetic modification, and hormone signaling. Among them, morphogenic factors have been extensively studied for enhancing transformation efficiency, while the potential of the other three pathways remains less explored in species beyond Arabidopsis. We summarize these established mechanisms and regulatory networks, providing a valuable reference for elucidating these mechanisms in other plant species. Furthermore, we also propose strategies for identifying species-specific efficient developmental factors and improving molecular mechanisms. Ultimately, this review provides a comprehensive summary of the mechanism and application of developmental factors and offers theoretical support to overcome the bottlenecks in genetic transformation.

Glyphosate-resistant crops, developed using the EPSPS gene, are widely cultivated globally. Previous studies indicate that the TIPS-OsEPSPS gene provides glyphosate resistance in rice. In this research, the OsmEPSPS gene was utilized to develop glyphosate-resistant, maize, and soybean, and a rapid immunochromatographic strip (ICS) test was designed for specific detection of the OsmEPSPS protein. The ICS test employs a double-antibody sandwich format with two monoclonal antibodies: mAb #9, conjugated to colloidal gold for capture, and mAb #15, for detection at the test line. The ICS exhibited a limit of detection (LOD) of 0.031 μg/mL within 10 min. Additionally, the ICS could detect a concentration of 0.817 μg/mL within 1 min in both Eleusine indica and OsmEPSPS-overexpressing plants. The ICS is highly specific, sensitive, and stable, with an R2 value of 0.982. Notably, the ICS is capable of detecting EPSPS proteins with high homology to OsmEPSPS from Poaceae and other plants. In summary, this study developed a sensitive and specific test strip for OsmEPSPS detection, facilitating semiquantitative analysis on the basis of colorimetric response time and intensity. This ICS is not only suitable for the rapid identification of OsmEPSPS-overexpressing plants but also for phylogenetic analysis of EPSPS gene homology.

Glyphosate resistance is a critically important trait for genetically modified (GM) crops. Mutation of the rice EPSPS gene results in a high level of glyphosate resistance, presenting significant potential for the development of glyphosate-tolerant crops. The resistance of rice to glyphosate is correlated with the expression levels of resistance genes. Therefore, developing a convenient, stable, and sensitive method for quantifying the OsmEPSPS protein is crucial for the development of glyphosate-resistant crops. We developed a double-antibody sandwich quantitative ELISA (DAS-ELISA) using a specific monoclonal antibody (mAb) for OsmEPSPS capture and an HRP-conjugated anti-OsmEPSPS rabbit polyclonal antibody (pAb). The method could be used to detect OsmEPSPS within a linear range of 16-256 ng/mL with robust intra- and inter-batch duplicability (%CV values: 0.17%-7.24%). OsmEPSPS expression in the transgenic rice lines (54.44-445.80 μg/g) was quantified using the DAS-ELISA. Furthermore, the expression of the OsmEPSPS gene was validated through Western blotting. This study demonstrated the reliability and stability of the DAS-ELISA for OsmEPSPS detection in GM rice.

This study analyzed the flavor compounds in the meat of four indigenous breeds of Chinese sheep through the use of gas chromatography–ion mobility spectrometry (GC-IMS). GC-IMS provided information on the characteristics and strength of 71 volatile flavor compounds (monomers and dimers), with aldehydes, alcohols and ketones being the most abundant in all types of sheep meat. The compounds with higher intensity peaks in the sheep meat were aldehydes (n-nonanal, octanal, heptanal, 3-methylbutanal, and hexanal), alcohols (1-octen-3-ol, hexanol, and pentanol), ketones (3-hydroxy-2-butanon, 2-butanone, and 2-propanone), esters (methyl benzoate), and thiazole (trimethylthiazole). The volatile flavor components in the meat of the different breeds of sheep obtained via GC-IMS were further differentiated using principal component analysis. In addition, orthogonal partial least squares discriminant analysis (OPLS-DA) and variable importance on projection (VIP) were used to determine the characteristic flavor compounds in the meats of different breeds of sheep, and 21 differentially volatile components were screened out based on having a VIP above 1. These results indicate that GC-IMS combined with multivariate analysis is a convenient and powerful method for characterizing and discriminating sheep meat.

Light intensity is a key factor affecting the synthesis of secondary metabolites in plants. However, the response mechanisms of metabolites and genes in Gentiana macrophylla under different light intensities have not been determined. In the present study, G. macrophylla seedlings were treated with LED light intensities of 15 µmol/m2/s (low light, LL), 90 µmol/m2/s (medium light, ML), and 200 µmol/m2/s (high light, HL), and leaves were collected on the 5th day for further investigation. A total of 2162 metabolites were detected, in which, the most abundant metabolites were identified as flavonoids, carbohydrates, terpenoids and amino acids. A total of 3313 and 613 differentially expressed genes (DEGs) were identified in the LL and HL groups compared with the ML group, respectively, mainly enriched in KEGG pathways such as carotenoid biosynthesis, carbon metabolism, glycolysis/gluconeogenesis, amino acids biosynthesis, plant MAPK pathway and plant hormone signaling. Besides, the transcription factors of GmMYB5 and GmbHLH20 were determined to be significantly correlated with loganic acid biosynthesis; the expression of photosystem-related enzyme genes was altered under different light intensities, regulating the expression of enzyme genes involved in the carotenoid, chlorophyll, glycolysis and amino acids pathway, then affecting their metabolic biosynthesis. As a result, low light inhibited photosynthesis, delayed glycolysis, thus, increased certain amino acids and decreased loganic acid production, while high light got an opposite trend. Our research contributed significantly to understand the molecular mechanism of light intensity in controlling metabolic accumulation in G. macrophylla.

MOTIVATION:Constructing a phylogenetic tree requires calculating the evolutionary distance between samples or species via large-scale resequencing data, a process that is both time-consuming and computationally demanding. Striking the right balance between accuracy and efficiency is a significant challenge. RESULTS:To address this, we introduce a new algorithm, MIKE (MinHash-based k-mer algorithm). This algorithm is designed for the swift calculation of the Jaccard coefficient directly from raw sequencing reads and enables the construction of phylogenetic trees based on the resultant Jaccard coefficient. Simulation results highlight the superior speed of MIKE compared to existing state-of-the-art methods. We used MIKE to reconstruct a phylogenetic tree, incorporating 238 yeast, 303 Zea, 141 Ficus, 67 Oryza, and 43 Saccharum spontaneum samples. MIKE demonstrated accurate performance across varying evolutionary scales, reproductive modes, and ploidy levels, proving itself as a powerful tool for phylogenetic tree construction. AVAILABILITY AND IMPLEMENTATION:MIKE is publicly available on Github at https://github.com/Argonum-Clever2/mike.git.

WUSCHEL (WUS) is a crucial transcription factor in regulating plant stem cell development, and its expression can also improve genetic transformation. However, the ectopic expression of WUS always causes pleiotropic effects during genetic transformation, making it important to understand the regulatory mechanisms underlying these phenomena. In our study, we found that the transient expression of the maize WUS ortholog ZmWus2 caused severe leaf necrosis in Nicotiana benthamiana. We performed transcriptomic and non-target metabolomic analyses on tobacco leaves during healthy to wilted states after ZmWus2 transient overexpression. Transcriptomic analysis revealed that ZmWus2 transformation caused active metabolism of inositol trisphosphate and glycerol-3-phosphate, while also upregulating plant hormone signaling and downregulating photosystem and protein folding pathways. Metabolomic analysis mainly identified changes in the synthesis of phenylpropanoid compounds and various lipid classes, including steroid synthesis. In addition, transcription factors such as ethylene-responsive factors (ERFs), the basic helix–loop–helix (bHLH) factors, and MYBs were found to be regulated by ZmWus2. By integrating these findings, we developed a WUS regulatory model that includes plant hormone accumulation, stress responses, lipid remodeling, and leaf necrosis. Our study sheds light on the mechanisms underlying WUS ectopic expression causing leaf necrosis and may inform the development of future genetic transformation strategies.

Ampelopsis grossedentata, commonly known as "Vine Tea" and well-recognized for its rich flavonoid content, is mainly distributed in the southern regions of the Yangtze River basin in China. These regions include Hunan, Hubei, Jiangxi, and Guizhou provinces. Vine Tea is mainly consumed as an herbal tea and has garnered attention for its reported health benefits, including antioxidant, anti-inflammatory, anti-tumor, anti-diabetic, and neuroprotective properties. It has been used to alleviate coughs and sore throats (Zhang et al., 2021; Wang et al., 2017; Gao et al., 2009). In the Zhangjiajie region of Hunan province alone, the Vine Tea planting area reached 7,670.5 hectares and produced commercial goods worth 1.417 billion RMB in 2022. In May 2021, leaf margins and veins fading to yellowing mottling, and crumpling of leaf blades in the shape of a boat symptoms were found in ~16% of Vine Tea plants in the Sanjiakuan Township, Yongding District, Zhangjiajie region (29°15'E, 110°30' N) (Figure 1a, b, c). (Figure 1a, b, c). Phytoplasma-like microbial cells (small oval shaped bacterial cells, around 1000 nm in size) were observed in sieve tube cells in the phloem of diseased leaves using transmission electron microscopy. No such cell was observed in the phloem of healthy leaves (Figure 2a, b). To investigate the potential association between phytoplasma and the observed symptoms of the diseased plants, total DNA was isolated from ten diseasedeaves and compared with ten healthy leaves from the same field using SteadyPure Plant Genomic DNA Extraction Kit. The isolated DNAs were analyzed first in a direct PCR using universal phytoplasma primer pair R16mF2/R16mR1 targeting the 16S rRNA gene (Gundersen and Lee 1996) and specific pair rpF1/rpR1 (Lee et al. 1998) targeting the DNA fragment encoding partial ribosomal proteins (rp) L22 (complete) and S3 and S19 (partial). The initial amplified products were used as templates and further amplified by nested PCR respectively with primer pair R16F2n/R16R2 for the 16S rRNA gene (Lee et al. 1998) and the rpF2/rpR2 primer pair for the rp gene (Martini et al. 2007). No amplification was obtained with DNA from healthy leaf samples using any of the four primer pairs. The amplified fragments from diseased leaves by nested PCR were cloned and sequenced (Qingke Biotech, China). The obtained sequences have been deposited in GenBank with accession numbers OR282806 for the 16S rRNA gene and GenBank OR353012 for the rp gene. BLASTn analysis revealed that the partial 16S rRNA gene sequence in our sample shared 99.4% nucleotide sequence identity with 'Candidatus Phytoplasma sp.' (MW364378) and 'Peony yellows phytoplasma' (KY814723) of the 16SrI group. Similarly, our rp gene sequence shared 99.6% nucleotide identity with the rpI group of phytoplasma such as the 'Balsamine virescence phytoplasma' (JN572890) and 'Paulownia witches'-broom phytoplasma' (HM146079). Phylogenetic analysis of the 16S rRNA and rp sequences using MEGA version 7.0 revealed that the phytoplasma strain associated with A. grossedentata yellow leaf syndrome in our study site belonged to the 16SrI (Candidatus Phytoplasma asteris) group of phytoplasma (Figure 3a, b). Using the interactive online phytoplasma classification tool iPhyClassifier (Zhao et al., 2009), virtual restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene sequences showed our strain having a distinct RFLP map but was closest to that of the onion yellow phytoplasma 16SrI-B subgroup (GenBank accession number: AP006628), with a similarity coefficient of 0.94 (Figure 4a, b). To confirm phytoplasma transmission, healthy plants were inoculated with three scions of infected plants of A. grossedentata. After 16 days, the new leaves of the inoculated A. grossedentata showed yellow leaf symptoms (Figure 5a, b, c), akin to the symptoms originally observed in the field, and the outer contour of the leaf margin appeared chlorotic. After 26 days, primer pairs R16mF2/R16R1 and R16F2n/R16R2 were used for nested PCR detection of phytoplasma in symptomatic A. grossedentata leaves. Phytoplasma was detected in the first and second leaves of symptomatic branches and leaves while negative control showed no amplification. Sequencing of the amplified fragments showed 100% nucleotide identity to the strain from the grafting source. Our results indicated that the pathogen and the disease can be transmitted by tissue grafting, consistent with the biological characteristics of phytoplasma, and further confirmed that the phytoplasma was the pathogen of yellow leaf syndrome of A. grossedentata. Toour knowledge, this is the first report of phytoplasma of group 16SrI affecting A. grossedentata.

Successful crop stand establishment is critical to realize high yield potential, which is dependent on depth of seed placement to access soil moisture. The coleoptile determines sowing depth by its length and ability to emerge from depth. This study was conducted to assess coleoptile length among three sets of three Great Plains winter small grain cereals—wheat ( Triticum aestivum L.), barley ( Hordeum vulgare L.) and triticale ( X triticosecale Wittm.)—and to evaluate the effect of the Rht-B1b dwarfing allele on coleoptile length in wheat and triticale. Fifty seeds of each genotype were sown in wet germination paper in two replications utilizing a randomized complete block design, which were placed in dark growth chambers at 25°C. Measurements were conducted after 7 d, and analysis of variance and comparison of least square means for coleoptile length among and within each set of genotypes were performed in Statistix 8.1 software using Fisher’s protected least significance difference at the α = 0.05 significance level. Results revealed that triticale had the longest coleoptiles, which were significantly longer ( P < 0.05) than those measured in both barley and wheat. Additionally, significant variation in coleoptile length ( P < 0.05) was also found within each set of wheat (3.52–6.41 cm), barley (4.32–6.63 cm) and triticale (4.05–6.92 cm) genotypes, respectively. These findings confirm other reports that the presence of the Rht-B1b allele was pleiotropic for coleoptile length, but development of semi-dwarf wheats with longer coleoptiles is possible if breeders deploy concurrent selection strategies.

Frozen storage can delay mutton meat spoilage to the maximum extent, prolonging its shelf life. In this experiment, mutton refrigerated for 24 h was used as the control group and that stored at -18 degrees C was used as the experimental group to explore the effects of frozen storage for 90 days on FA, AA, and VOC of the mutton. The results showed that FA, AA, and VOC changed significantly during storage. PUFA and n-3 FA contents were the highest at 3 days of freezing (54.92 and 15.28 mg/100 g, respectively). AA contents were higher in the experimental group than in the control group, and Glu contents were the highest during the whole storage period, ranging from 2.94% to 3.28%, and the maximum value was on the 3rd day. In total, 38 volatile compounds were detected, and the most abundant volatile compounds were detected at 60 days. The variance contribution rates of PCA were 48.7%, 19.4%, and 14.8%, the PCA model can be used as a separation model. Novelty impact statement This study simulated the storage of mutton by household users and shortened the detection time point at the initial freezing stage, indicating that the differences in flavor substance changes in mutton at the initial freezing stage was little. On the other hand, GC-IMS was effectively used to visualize flavor substance changes during storage, providing a new method to distinguish the storage time of meat products. In conclusion, this study can provide basic research data for long-term storage of mutton and provide technical reference for meat storage identification.

Low propagation rate is the primary problem that limits industry development of tree peony. In this study, a highly efficient regeneration system for tree peony using somatic embryogenesis (SE) was established. The transcriptomes of zygotic embryo explants (S0), non-embryonic callus (S1), embryonic callus (S2), somatic embryos (S3), and regenerated shoots (S4) were analyzed to determine the regulatory mechanisms that underlie SE in tree peony. The differentially expressed genes (DEGs) were identified in the pairwise comparisons of S1-vs-S2 and S1-vs-S3, respectively. The enriched DEGs were primarily involved in hormone signal transduction, stress response and the nucleus (epigenetic modifications). The results indicated that cell division, particularly asymmetric cell division, was enhanced in S3. Moreover, the genes implicated in cell fate determination played central roles in S3. Hormone signal pathways work in concert with epigenetic modifications and stress responses to regulate SE. SERK, WOX9, BBM, FUS3, CUC, and WUS were characterized as the molecular markers for tree peony SE. To our knowledge, this is the first study of the SE of tree peony using transcriptome sequencing. These results will improve our understanding of the molecular mechanisms that underly SE in tree peony and will benefit the propagation and genetic engineering of this plant.

离子迁移谱(ion mobility spectrometry,IMS)是一种利用样品分子电离后离子迁移率的差异进行分离检测的仪器.它具有灵敏度高、选择性好、分析灵活、携带方便、实时监测能力强等优点.因此,广泛应用危险品检测、化工分析、食品检测、医学诊断、生物制药研究及环境监测等领域.该文简要介绍离子迁移谱技术的工作原理、分类及联用技术,综述IMS技术在食品真伪鉴别、加工储藏过程监测、非法添加物识别和有害物质检测等方面的应用,并对IMS在食品检测领域的进一步发展进行展望.

In this study, gas chromatography coupled to an ion mobility spectrometry (GC-IMS) was used for analyzing some volatile components and flavor fingerprint in samples from Jingyuan lambs of different ages (2, 6, and 12 months). The data obtained from ion mobility were processed using laboratory analysis view processing software for fingerprint recognition, and the principal component analysis (PCA) was performed. GC-IMS provided information on the characteristics and strength of 66 volatile flavor compounds (monomers and dimers). The differences in flavoring substances between lambs of different ages were observed. The compounds with higher intensity peaks in the lamb meat samples were alcohols (1-octen-3-ol, ethanol, (E)-2-hexen-1-ol, 1-pentanol, and 2-propanol), ketones (2-pentanone, 2-heptanone, 3-hydroxy-2-butanone, 2-hexanone, 2-butanone, 2-propanone, and 4-methyl-2-pentanone), aldehydes (n-nonanal, octanal, heptanal, 3-methylbutanal, hexanal, pentanal, 2-methylbutanal, (E)-2-octenal, (E)-2-nonenal, methional, and phenylacetaldehyde), esters (methyl benzoate), furan (2-pentylfuran), and thiazole (trimethylthiazole). The results showed that the flavor fingerprint in samples from Jingyuan lambs of different ages (2, 6, and 12 months) can be established by GC-IMS and PCA based on the identified volatile compounds. This method might be used for the rapid and comprehensive analysis of volatile components in lamb meat.

选择6月龄滩羊股二头肌、背最长肌和臂三头肌3个部位肌肉为实验材料,进行挥发性化合物和脂肪酸的测定.采用气相色谱-离子迁移谱技术分析滩羊肉中的挥发性风味物质,并建立指纹图谱.结果 表明:在滩羊肉中共鉴定出44种挥发性风味物质,包括醛类15种、醇类11种、酮类9种、酯类6种、噻唑类2种和呋喃类1种,主要风味化合物为己醛、壬醛、1-辛烯-3-醇、3-羟基-2-丁酮和乙酸乙酯.依据所建立的指纹图谱信息,采用主成分分析法对不同部位羊肉样品进行分析,主成分1和主成分2的贡献率之和达到了90％,3个部位羊肉样本特征风味物质具有明显的差异.滩羊肉中背最长肌、股二头肌和臂三头肌中饱和脂肪酸/不饱和脂肪酸分别为0.84、0.92、0.93,均接近于营养协会推荐人体摄入脂肪酸的组成比例1∶1.其中,背最长肌中的多不饱和脂肪酸含量显著高于臂三头肌,丰富的不饱和脂肪酸含量,对羊肉风味具有十分重要的作用.

豌豆(Pisum sativum L.)是一种重要的食用豆类作物,在全世界范围内广泛种植,既可作为人类食物,也可作为牲畜饲料.用SSR标记构建的遗传连锁图谱在豌豆和其他作物的标记辅助育种中发挥着重要的作用.尽管对豌豆遗传连锁作图的研究已有悠久历史,但公众可获得且可转移的SSR标记以及基于遗传独特的中国豌豆种质的高密度遗传连锁图谱仍然有限.为了获得更多可转移的SSR标记和中国豌豆的高密度遗传连锁图谱,本研究首先从自主开发和文献获取的12,491个全基因组SSR标记中筛选了617个多态性SSR标记,并用于G0003973×G0005527 F2群体遗传连锁图谱的加密.加密后的图谱全长扩展到5330.6 cM,包含603个SSR标记,标记平均间距离8.8 cM,相比之前的图谱有明显改善.基于上述结果,我们又筛选了119个具有多态性的SSR标记,用于构建大样本W6-22600×W6-15174 F2群体的遗传连锁图谱,新图谱累积长度为1127.1 cM,包含118个SSR标记,装配在7条连锁群上.最后,将来自以上2个遗传图谱的数据进行整合,得到了一张覆盖范围6592.6 cM的整合图谱,包含668个SSR标记,由509个基因组SSR、134个EST-SSR和25个锚定标记组成,分布在7条连锁群上.这些SSR标记和遗传连锁图谱将为豌豆的遗传研究和标记辅助育种提供有力工具.

[Objective]Millet planting area is hilly and mountainous terrain in Shanxi Province,the seeding efficiency is low,to increase the seeding efficiency,[Methods]according to the topographic features of millet planting area in Shanxi province,with millet free thinning precision seeding agronomic requirements,we designed a millet precision seeding machine using human and animal power.In this article,the main structure parameters of seeding machine was calculated,and its working process was expounded.We did test on seeding performance,the indexes of test were variation coefficient of seeding stability,variation coefficient of evenness of seeding rate between rows,variation coefficient of sowing uniformity and seed damage rate.[Results]The results showed that the variation coefficient of seeding stability was less than 2.5 ％,the variation coefficient of evenness of seeding rate between rows was less than 2.7 ％,the variation coefficient of sowing uniformity was less than 41 ％,the seed damage rate was less than 0.5 ％.It was adapted to the national industry standards.[Conclution]The design of precision seeding machine using human and animal power for millet was to provide a basis of the manufacture seeding machine for Shanxi terrain features.

Pea (Pisum sativum L.) is an important food legume globally, and is the plant species that J.G. Mendel used to lay the foundation of modern genetics. However, genomics resources of pea are limited comparing to other crop species. Application of marker assisted selection (MAS) in pea breeding has lagged behind many other crops. Development of a large number of novel and reliable SSR (simple sequence repeat) or microsatellite markers will help both basic and applied genomics research of this crop. The Illumina HiSeq 2500 System was used to uncover 8,899 putative SSR containing sequences, and 3,275 non-redundant primers were designed to amplify these SSRs. Among the 1,644 SSRs that were randomly selected for primer validation, 841 yielded reliable amplifications of detectable polymorphisms among 24 genotypes of cultivated pea (Pisum sativum L.) and wild relatives (P. fulvum Sm.) originated from diverse geographical locations. The dataset indicated that the allele number per locus ranged from 2 to 10, and that the polymorphism information content (PIC) ranged from 0.08 to 0.82 with an average of 0.38. These 1,644 novel SSR markers were also tested for polymorphism between genotypes G0003973 and G0005527. Finally, 33 polymorphic SSR markers were anchored on the genetic linkage map of G0003973 × G0005527 F2 population.

The study of genetic diversity between Lathyrus sativus L. and its relative species may yield fundamental insights into evolutionary history and provide options to meet the challenge of climate changes. 30 SSR loci were employed to assess the genetic diversity and population structure of 283 individuals from wild and domesticated populations from Africa, Europe, Asia and ICARDA. The allele number per loci ranged from 3 to 14. The average gene diversity index and average polymorphism information content (PIC) was 0.5340 and 0.4817, respectively. A model based population structure analysis divided the germplasm resources into three subgroups: the relative species, the grasspea from Asia, and the grasspea from Europe and Africa. The UPGMA dendrogram and PCA cluster also demonstrated that Asian group was convincingly separated from the other group. The AMOVA result showed that the cultivated species was quite distinct from its relative species, however a low level of differentiation was revealed among their geographic origins. In all, these results provided a molecular basis for understanding genetic diversity of L. sativus and its relatives.

Simple sequence repeat(SSR)markers have previously been applied to linkage mapping of the pea(Pisum sativum L.)genome.However,the transferability of existing loci to the molecularly distinct Chinese winter pea gene pool was limited.A novel set of pea SSR markers was accordingly developed.Together with existing SSR sequences,the genome of the G0003973(winter hardy)×G0005527(cold sensitive)cross was mapped using 190 F2individuals.In total,157 SSR markers were placed in 11 linkage groups with an average interval of 9.7 cM and total coverage of 1518 cM.The novel markers and genetic linkage map will be useful for marker-assisted pea breeding.