Abstract:EMS is the most commonly used mutagen in chemical mutagenesis， which has the advantages of high point mutation rate of a single base， low cost and easy operation. Obtaining mutants through EMS mutagenesis can provide favorable material for breeding and gene function studies. The key of EMS mutagenesis technology is to determine the EMS concentration and mutagenesis time. Generally， the combination of concentration and time to achieve the half lethal rate is the best. In gramineous plants， seeds are mainly used as mutagenic materials. At the same time， pollen， callus and vegetative organs of gramineous plants that rely on vegetative reproduction can be used as mutagenic materials. Different plant materials have different tolerance to EMS. Pollen is the most sensitive， followed by callus， and asexual reproduction materials and seeds have strong tolerance. The screening methods of mutants include phenotypic comparison screening， stress directional screening， and forward and reverse genetic screening. This article reviews the application of EMS mutagenesis in gramineous plants breeding and gene function research in recent years， introduces the mutagenesis principle， mutagen concentration， mutagen time， mutagenic material selection and the screening of mutants， and the future research on EMS mutagenesis in gramineous plants was prospected， this information can provide reference for EMS mutagenesis research of gramineous plants in the future.

Abstract:With the global climate change， the probability of extreme weather events has significantly increased. As an important base for commercial rice production in China， Heilongjiang province is an area prone to low-temperature stresses. Geng/Japonica rice in Heilongjiang is susceptible to low-temperature stresses during both the seedling stage and the reproductive stage （including booting， flowering， and maturing stages）. Low temperature stress is a key limiting factor for rice production in Heilongjiang， and improving cold tolerances of Heilongjiang Geng/japonica rice cultivars is of great strategic importance for ensuring the food production security in China. When reviewing the identification methods and genetic researches of rice cold tolerance， the authors analyzed the cold tolerance characteristics of Heilongjiang rice cultivars released in the past about 20 years （2006-2023）， and found that with the "blowout" of approved cultivars in recent 5 years， the cold tolerances are going down. Secondly， through the comparative mapping based on reference genome， it was found that the cold tolerances at the seedling stage and the reproductive stage are mostly controlled by independent loci or chromosomal regions， and the proportion of genetic overlap （including both pleiotropic loci and linked regions） accounted for about 21%. Among the identified gene responsible for cold tolerances， the negative regulatory genes accounted for about 20%. For current breeding application， the above-mentioned genetic overlap loci/regions and negative regulatory genes are useful in improving the breeding efficiency. On this basis， the authors put forward specific suggestions on the simultaneous improvement of cold tolerance during the seedling stage and reproductive stage as well as strategies for the biotechnological breeding on improving the cold tolerances of early Geng/japonica rice for Heilongjiang province.

Abstract:In view of the increasingly serious problem of the endangered status of wild rice in Guangxi， in order to rescue the germplasm resources of wild rice in Guangxi and explore its disease resistance genes， the areas with abundant wild rice resources in Guangxi were investigated and collected， and the resistance of white leaf blight and rice blast was identified in some of the collected resources. The results showed that only 24 of the 44 distribution points recorded in the wild rice field in Guangxi still had wild rice distribution， and the endangered situation was very serious， and the proportion of wild rice distribution points disappeared as high as 45.5% in the past 10 years， and the destruction of water sources and weed competition were the main threats to the survival of wild rice. According to the principle of wild rice sampling， a total of 317 germplasm resources from 11 groups of Oryza rufipogon Griff. and Oryza officinalis Wall. were rescued； The resistance to bacterial blight was identified by using the international virulent strain PXO99， and the resistance to rice blast of different isolates （ Guy11， RB22， FJ-3-2， FJ-3-5， FJ-2-3 ） was identified by wounding detached rice leaf. The results showed that 27 bacterial blight resistance resources， 105 rice blast resistance resources， and 5 resources with both bacterial blight resistance and rice blast resistance were identified from 177 wild rice. In view of the problem that the disease-resistant resources of wild rice in Guangxi are rich but disappear seriously， it is suggested to strengthen the collection and protection， and increase the identification of germplasm resources.

Abstract:The objective of this study was to investigate the correlation between the quality traits of peanut germplasm resources from different sources and provide a theoretical basis for the rational exploration of fresh peanut germplasm resources. We utilized genetic diversity analysis， correlation analysis， principal component analysis ， clustering analysis， and comprehensive score to analyze and assess the 11 quality traits of 287 peanut germplasm resources.The results showed that the coefficient of variation of 11 quality traits ranged from 1.286% to 19.506%， and the genetic diversity index ranged from 1.046 to 2.073. The results of correlation analysis showed that the oleic acid content has an extremely significant negative correlation with proteins content and an extremely significant positive correlation with sucrose content. A total of three principal component factors were extracted from the principal component analysis， and their cumulative contribution rate reached 71.467%. Cluster analysis divided the 287 materials into 3 groups.The first group has a higher content of fat and stearic acid，which contains 100 materials；the second group has a higher oleic acid content ，which contains 61 materials；and the third group has the characteristics of high protein content and low fat content，which contains 126 materials. By assigning weights of the contribution rate of the 3 principal components， we constructed a comprehensive scoring formula： F=0.588F₁+0.277F₂+0.135F₃. According to this formula， we selected 51 materials with a comprehensive score greater than 1， including 3 materials with a score exceeding 5.This research provides valuable insights for future studies in peanut quality breeding.

Abstract:The genetic diversity analysis and comprehensive evaluation of 686 safflower germplasm accessions were conducted via investigating the phenotypic variations， followed by the coefficient of variation， correlation analysis， principal component analysis， and cluster analysis， aiming to provide a foundation for safflower breeding in Xinjiang， P.R. China. The coefficient of variation of 11 agronomic characters ranged from 12.72% to 44.80%， indicating rich variations at these traits in the collection. The genetic diversity index of the three quality traits ranged from 0.75 to 1.04. Correlation analysis showed that yield per plant was significantly positively correlated with plant height， number of primary branches， number of secondary branches， individual bulb number， head ball diameter and total number of per fruit. When the genetic distance of 7.5 applied， the germplasm resources were clustered into four groups， in which the yield per plant and thousand grain weight of group IV were excellent. Principal component analysis showed that the cumulative contribution rate of the five principal component factors was 78.838%. Among them， leaf margin， leaf spines， individual bulb number， total number of per fruit， and thousand grain weight were the main factors contributing to the phenotypic differences of safflower. Three elite germplasm Hefeihonghua （220657）， 78-1 （220675） and Linquanhonghua （220683） were identified by comprehensive evaluation， among which Linquanhonghua （220683） showed a higher 1000-grain weight （53.42g）， serving as parental line in high-yield breeding. Collectively， this study showed rich genetic diversity of 686 safflower germplasm resources， which could provide basic materials in breeding for new safflower varieties.

Abstract:In order to evaluate salt tolerance in sunflower germplasm resources and identify elite accessions， the salt tolerance of 444 sunflower germplasm resources was evaluated under 250 mmol/L NaCl stress at germination and seedling stage， and also assessed under saline-alkali soil condition at the whole growth period.The salt tolerance analysis and evaluation was performed by observing a number of traits including the relative values（ratio of each indicator compared to the control treatment）of germination rate at germination stage，six indexes at seedling stage （survival rate，leaf area，plant height，SPAD value，etc.）and seven indexes at the whole growth period（plant height，sunflower diameter，seed setting rate，yield related traits，etc.） The correlation analysis results indicate that there was a highly significant positive correlation between each two indexes at seedling stage， with correlation coefficients ranging from 0.518-0.790. The correlation between most of the seven indicators in the whole growth period reached a significant or extremely significant level. The analysis of principal component， membership function value， cluster and comprehensive salt tolerance evaluation were performed based on these relative indicator values， then 132 salt resistant materials at germination stage， 9 at seedling stage， and 41 at whole growth stage were identified. Six germplasm resources （ZX0365， ZX0389， ZX1391， ZX1394， ZX3089， ZX3094）， which showed salt tolerance in the three periods， can be utilized for subsequent salt-tolerant gene mining and breeding. Correlation analysis of salt tolerance levels for 124 accessions at three stages （germination， seedling， and the whole growth period） indicated a highly significant correlation among the salt tolerance assessments at each stage. The salt tolerance assessments of the germination and seedling stages can provide effective references for selecting salt-tolerant materials when planting sunflower in saline-alkali soil.This study provided method and material basis for the selection of salt-tolerant sunflower varieties.

Abstract:This study aims to identify the pre-harvest sprouting resistance in wheat germplasm resources and molecular markers for marker-assisted selection， ultimately obtaining excellent white grain wheat germplasm resources applicable in wheat breeding. These spikes of 222 wheat germplasm resources were tested for pre-harvest sprouting resistance， and the functional markers of eight anti-pre-harvest sprouting genes， including myb10D， DFR-B， Vp1B3， PM19-A1， MFT-3A， MFT-A2， MKK3-A， QSD1， were used for genotyping. Phenotypic identification results showed that there were significant differences in the relative sprouting percentage of 222 wheat germplasm resources. The relative sprouting percentage ranged from 0 to 1.15， and the average relative sprouting percentage was 0.73. 38 wheat materials were identified to be resistant to pre-harvest sprouting， including 9 white wheat， 27 red wheat and 2 black wheat. Correlation analysis between allele types and relative sprouting percentage showed that the relative sprouting percentage was significantly correlated with functional markers myb10D， DFR-B， Vp1B3， MFT-3A and MFT-A2， but not with PM19-A1， MKK3-A and QSD1. These results indicated that molecular markers myb10D， DFR-B， Vp1B3， MFT-3A and MFT-A2 could be used for detection of pre-harvest sprouting resistance and marker-assisted breeding. Based on the phenotype and genotype results， nine white wheat germplasm resources with pre-harvest sprouting resistance were selected， including Yunong 914， Yunong 946， Fengdecunmai 30， Fangmai 5， Xumai 029， Lianmai 1901， Baofeng 1903， Zhengmai 829 and 13wang27-8， which could be used for genetic breeding and layout of wheat varieties with pre-harvest sprouting resistance.

Abstract:Molecular markers are the basic tools of genetic research， which are widely used in genetic diversity research， germplasm identification， genetic map construction and gene mapping. In this study， Barley SNP 50K chip was used to detect SNP in barley core resources with wide genetic background， and a series of dimorphic SNPs with high polymorphism were selected， and 124 KASP markers were developed. 43 barley germplasms from different geographical sources were used to test their effectiveness， and 56 KASP markers were initially screened out. Eighteen high-quality KASP markers were selected based on MAF （minor allele frequency）≥0.40 and PIC （polymorphic information content）≥0.45 criteria and applied to construct phylogenetic trees of 98 Jiangsu barley varieties. The results showed that barley materials with the same geographical origin and close relatives could be grouped into one class. These results indicate that KASP markers have guiding significance and application value in barley variety identification genetic relationship analysis of barley resources and population division. At the same time， the SNP fingerprint of 98 Jiangsu barley varieties was constructed， which verified the feasibility of KASP technique in barley varieties identification in China. The developed KASP marker can identify barley varieties accurately and quickly ， which is helpful to the scientific and standardized management of barley germplasm resources and the study of genetic diversity.

Abstract:By mining QTL significantly associated with zinc content in rice grains， combined with gene function annotation， haplotype and grain relative expression analysis at grain filling stage， candidate genes related to zinc content in rice grains were screened. In this study， 139 core germplasm resources of early japonica rice in Northwest China were used as experimental materials. Combined with 1800920 high-confidence SNPs obtained by resequencing， genome-wide association analysis was performed based on the general linear model （GLM）. The candidate genes in the significant QTL interval were functionally annotated， and the further screened genes were analyzed for haplotypes and relative expression levels at grain filling stage， in order to screen out candidate genes and dominant haplotypes related to grain zinc content. The results showed that there were abundant phenotypic variations in grain zinc content of 139 rice accessions， which were in accordance with normal distribution. Through genome-wide association analysis of zinc content in rice grains， a total of 12 significant SNPs were associated， which were distributed on chromosomes 1， 2， 4， 10 and 11. Combined with the gene function annotation in the candidate interval， it was speculated that OsFRDL2 located in the vesicle may be related to the enrichment of zinc in rice grains. Through the analysis of haplotype and grain relative expression at filling stage， it was found that OsFRDL2 was relatively highly expressed in the dominant haplotype Hap5. Therefore， it is speculated that OsFRDL2 may be related to the enrichment of zinc in rice grains. The results provide a theoretical basis for analyzing the mechanism of zinc enrichment in rice grains and developing molecular markers of zinc-rich rice， and provide genetic resources for cultivating zinc-rich rice.

Abstract:The type-B authentic response regulator （B-ARR） family members are positive regulators in cytokinin signal transduction， and play important roles in plant growth and development and resistance to abiotic stresses. However， there are few studies on the B-ARR gene family in wheat. In this study， 25 B-ARR gene family members were identified from wheat genome， and their physicochemical properties， gene structure， cis-acting elements and abiotic stress-induced expression patterns were analyzed by bioinformatics methods. The results showed that all B-ARR proteins were localized in the nucleus based on bioinformatics prediction， and their secondary structure was mainly consisting of α-helix and random crimp. B-ARR genes were not evenly distributed on wheat chromosomes， and the number of B-ARR genes was the highest on chromosome 7. In addition， multiple cis-acting regulatory elements related to growth and development， hormone response， and biological and abiotic stress have been identified in the promoter regions. RT-qPCR analysis showed that the relative expression of TaARRM-like9， TaARRM-like10， TaARRM-like12 and TaARRM-like13 were significantly up-regulated under abiotic stresses treatments， including drought， salt and low temperature. This study laid a foundation for further research on the role of B-ARR transcription factor in wheat development and abiotic stress response.

Abstract:Nitrogen content in maize grains is closely related to maize quality， and the analysis of its genetic mechanism is great significance for maize quality breeding. In this study， we used 252 maize inbred lines as an association population， and used bayesian-information and linkage-disequilibrium iteratively nested keyway （BLINK）， fixed and random model circulating probability unification （FarmCPU）， general linear model （GLM）， mixed linear model （MLM）， multiple loci mixed model （MLMM）， and settlement of MLMs under progressively exclusive relationship （SUPER） to conduct genome-wide association analysis for grain nitrogen content. A total of thirteen SNPS （P<3.64E-07） were identified. Six， three， seven， four， two and four SNPs were detected by BLINK， FarmCPU， GLM， MLM， MLMM and SUPER methods， respectively. S3_ 8879213 can be detected in five methods， S9_146173702 can be detected in four methods， S5_114774030 and S7_ 182217338 can be detected in three methods， S1_10906326， and S1_177528813 can be detected in two methods. A total of twenty-five candidate genes were identified， among which Zm00001eb275080 and Zm00001eb330700 may be the important candidate genes affecting maize grain nitrogen content.

Abstract:The heading date of foxtail millet is a key trait that determined the adaptability of varieties， and dissection of the transcriptional pattern and haplotype variations of key genes are important for promoting cultivars improvement. In this research， SiGI was identified by genome-wide association study （GWAS）， which is a key regulator responsible for foxtail millet flowering. The expression profile of SiGI was analyzed， as well as subcellular localization of SiGI was performed by using protoplasts. The 24-hour rhythm expression pattern analysis of SiGI was performed by qRT-PCR under short-day （10 h light/14 h dark） conditions. Genetic and haplotypes diversity of SiGI coding and promoter regions were analyzed by using representative 697 millet varieties， and haplotype morphological effects of SiGI was also summarized. As a result， a significant correlation signal was identified at 11062649 bp on chromosome 5， which is closely related to heading date. SiGI was found near this locus， whose homologue is AtGI. SiGI was highly expressed in photoperiod-responsive tissues （roots， stems， leaves， etc.）， and subcellular localization was located in the nucleus， and the expression level was up-regulated in the evening， showing a 24-hour rhythmic expression pattern. SiGI exhibits rich polymorphism in different foxtail millet varieties， and the relative expression of promoter haplotype Hap-6 was significantly upregulation by approximately 1.5-flod compared to Hap-3 （P=0.0083）. Heading date of Hap-6 containing varieties were significantly earlier than other haplotypes under 8 environments， and the plant height of Hap-6 containing varieties were significantly reduced under 4 environments. SiGI haplotype Hap-6 has no obvious impact on yield， and can be used as the main haplotype for molecular breeding selection.

Abstract:Sorghum plays an important role in agricultural farming and brewing industry of China. The grain color is one of the important traits， while its genetic basis remains largely unclear. By observing the grain development process using yellow grain material GLB41， we found that on 1 to 16 days after flowering the grain grew rapidly， followed by the slow expansion on 17~24 days， and the coloring on 25 days after flowering. At the coloring stage， the green color gradually faded， the grain color gradually changed from milky white or pale white to light yellow， and then the color deepened， and the grain color changed to dark yellow after 40 days. The genetic locus controlling the yellow grain trait was initially mapped to a 15.6 Mb interval on chromosome 1， and was further delimited between two markers BR13 and P2 by fine mapping in 3215 individuals. The functional annotation suggested seven candidate genes. Through Sanger sequencing of the candidate genes in GLB41 （yellow grain） and 6E16 （white grain）， we found three sequence variations （insertion at 619’- 621’， CTG/Leu； mutation at 819’， Cys to Trp； synonymous mutation at 912’， C to T） at the unreported gene Yellow Seed 2 （Sobic.001G397900）.Therefore， it is speculated that Yellow Seed 2 may be involved in the formation of the grain color of these two parent materials， as a new gene for further deciphering the functional mechanism of grain color in sorghum.

Abstract:Chitinase is a kind of glycosyl hydrolases （GH） which hydrolyzes the chitin and other polymers. Chitinase plays an important function in the plant growth and development， as well as in the resistant process to diverse stresses. However， the tissue expression patterns and responses to diverse stresses of chitinase genes in soybean are still unclear， which seriously limited its application in genetic improvement. In this study， the chitinase family genes were identified in the wild soybean （Glycine soja Sieb. and Zucc.） and cultivated soybean （Glycine max（L.）Merr.）， and the expression patterns were also analyzed. The results showed that 62 and 55 chitinase genes were identified in the wild soybean and cultivated soybean， which located on 17 and 18 chromosomes， respectively. The phylogenetic tree analysis showed that the chitinase genes were divided into five categories， with Class III and Class V belonging to the GH18 subfamily， while Class I， Class II and Class IV belonging to the GH19 subfamily. Further analysis found many cis-acting elements in the promoter regions of chitinase genes responding to various plant hormones and stresses.Further gene expression analyses in cultivated soybean showed that the chitinase genes presented differential expressions in different tissues and under different stress conditions. Among these genes， Gm.01G142400 and Gm.13G346700 were strongly induced in the leaves of resistant variety after soybean mosaic virus inoculation， Gm.03G254300 and Gm.20G164600 were induced in soybean roots after low phosphorus treatment， and Gm.08G259200 and Gm.19G245400 were induced in soybean nodules under low phosphorus condition. Gene expression analysis in wild soybean showed that the chitinase genes presented differential expressions in different tissues and after salt stress， among which Gs.02G002604 and Gs.02G002940 were highly induced in the leaves of tolerant variety after salt treatment. These results provide important references for further utilizing the chitinase genes in soybean genetic improvement.

Abstract:Polianthes tuberose L. is a perennial bulbous flower of the Amaryllidaceae family. It is a famous fragrant flower plant with high ornamental， economic and medicinal values. Bulbs are their main reproductive and medicinal organs. In order to further explore its growth and development mechanism and secondary metabolic pathways， we use the high-throughput sequencing PacBio Sequel platform to make use of its bulbs as materials， the single molecule long reading sequencing technology （SMART） was used to sequence and analyze the full-length transcriptome of Agave amica bulbs for three generations. The platform obtained a total of 508012 high-quality CCS reads and 402032 full-length non chimeric sequences （FLNC）， and clustered the full-length non chimeric sequences to obtain a total of 137215 high-quality consistent sequences. After quality control and redundancy removal， a total of 81719 transcripts （Unigene） and 36215 complete CDS regions were obtained. All transcripts obtained were annotated and functionally classified through databases such as NR， SwissProt， KOG，KEGG GO， etc. The result showed that a total of 64362 single genes were annotated， with NR having the highest annotation count of 63538， accounting for 98.72%，while KEGG had the lowest number of annotated genes，with 58878 genes annotated to 148 pathways. And 41240 SSRs， 15835 lncRNAs and 4903 transcription factors were analyzed. RLK transcription factor family has the most genes. The discovery of homologous box transcription factors family may be involved in the development process of tuber， PtKNOX1L13 gene was cloned，its expression levels were relatively high during the initiation， enlargement， and maturity stages of tubers， transient overexpression indicates that it can promote the development of bulbils，so that it may be an important functional gene involved in bulb formation and development.This study provides reference and basis for in-depth research and development of molecular markers， important functional genes and germplasm resources innovation of Polianthes tuberose L.

Abstract:The impact of climate change on the ecological suitability of species is a crucial concern for biodiversity conservation. This study employed a combination of 128 reliable distribution records of Paris polyphylla var. yunnanensis in China and 32 bioclimatic variables utilizing the MaxEnt model and ArcGIS software， in order to investigate the primary ecological factors that influence the distribution of ecological suitability for this species. Furthermore， we have made projections regarding the spatial distribution pattern of potential suitable areas and the trajectory of centroid displacement during the last glacial maximum， the middle holocene， the present， and future periods （2050s， 2070s） under three distinct greenhouse gas emission scenarios （RCP2.6， RCP4.5， RCP8.5）. The findings indicate that the MaxEnt model exhibited a remarkable level of precision in its predictions， as evidenced by a mean AUC value of 0.951 across all time periods. Notably， the geographical distribution of the studied entity was primarily influenced by several significant bioclimatic factors， namely precipitation annually， seasonality of temperature， warmest quarter precipitation， and elevation. Since the last glacial maximum， the areas of significant change in the potential habitat of Dianzhilou have been concentrated in the high habitability zone. Particularly， under the projected scenario of high greenhouse gas emissions in the future. In this scenario， the area of shrinkage reaches a maximum of 16.86×10⁴ km²， primarily concentrated in the northeastern region of Sichuan province. Simultaneously， the analysis of centroids revealed a tendency for the distribution of P. polyphylla var. yunnanensis to shift towards latitudes and altitudes of greater magnitude in response to climate change scenarios. These findings suggested that global warming contributed to the contraction of areas highly conducive for P. polyphylla var. yunnanensis. Consequently， the outcomes of this study furnish a theoretical framework for the judicious utilization of wild resources pertaining to P. polyphylla var. yunnanensis.