Photosynthesis is closely related to crop yield. Fast chlorophyll fluorescence parameters can sensitively reflect the electron transport rate in photosynthesis. Studying the genetic basis of fast chlorophyll fluorescence parameters can help in breeding maize (Zea mays) hybrid with high photosynthetic efficiency. In this study, 160 recombinant inbred lines (RILs) derived from maize hybrid 'Xianyu335' by continuous self-crossing for 7 generations were used to construct a high-density genetic map and map QTLs for fast chlorophyll fluorescence parameters. For this purpose, 5 parameters including ABS/CS_o, TR_o/CS_o, ET_o/CS_o, ET_o/TR_o, PIcs were measured in the RILs grown under 4 field conditions. A high-density SNP genetic map with a total length of 1 896.423 cM was constructed by SNP molecular markers originated from 20K molecular chips. The genetic map contained 2 915 markers, and the average distance between markers was 0.66 cM. A total of 15 related QTLs were identified using the best linear unbiased predictor (BLUP) of fast chlorophyll fluorescence parameters in multiple environments. The logarithm of odds (LOD) value were 2.52~7.35 and the explicable phenotypic variation were 4.56%~14.81%. Three of these QTLs were stably expressed in multiple environments, namely qREC3-1, qRPI2-1, and qRPI3-2. This study could provide a reference for further fine mapping of QTLs of rapid chlorophyll fluorescence parameters and molecular marker-assisted selection for high photosynthetic efficiency breeding.

Water-soluble carbohydrate (WSC) reserved in wheat stems is one of the important carbon sources for grain filling, and its inheritance is controlled by polygenes. It is an effective way for wheat molecular genetic improvement to explore the major QTL underlying WSC content through meta-analysis. In this study the bioinformatics strategy was used to integrate, and meta-analyze the previously reported 239 QTLs that involved in stem WSC content from different genetic populations in wheat, based on a high-density genetic map as a reference map. A consensus genetic map for stem WSC QTL was established, and a total of 39 meta QTLs (MQTL) which has the minimum confidence interval of 0.50 cM.These MQTLs were mainly distributed on chromosomes 1A, 2B, 2D, 3A, 3B, 4A, 4B, 6A, 6B, 7B and 7D, each of which carried 1 to 2 QTL clusters. In the marker interval of Eps-1Am~Xgwm99 on chromosome 1A, seven candidate genes related to stem WSC content were predicted. This study provides a reference for further shortening the confidence interval QTL wheat stem WSC content and QTL molecular marker-assisted selection breeding.

It has been more than 20 years since wheat (Triticum aestivum) molecular breeding was introduced to China, only a few commercial cultivars were released. To promote combination of marker assistant selection and traditional breeding, and accelerate molecular breeding development is urgent. There are 4 well characterized pleiotropic disease resistance genes in wheat. It is difficult to select these genes in breeding program through phenotype, marker assistant selection is an option. Markers developed for pleiotropic disease resistance genes have some shortcoming, such as ambiguous genotyping or single marker type, which limit its use. In this study, Focus on the ambiguous genotyping problem of reported kompetitive allele-specific PCR (KASP) markers of pleiotropic disease resistance genes, the basic local alignment search tool (BLAST) was used to align the KASP marker amplification regions and the 'Chinese Spring' reference genome. Except the target regions, several other similar genomic regions were found. Too many similar sequences in the genome may be the reason for genotyping problem of KASP markers. According to the BLAST result, primers were redesigned. The KASP marker genotyping result of improved Lr34K2 and Lr46K3 were better than Lr34_TCCIND for leaf rust resistance gene 34 (Lr34) and Lr46_JF2-2 for Lr46, respectively; KASP markers Sr2K3 for stem rust resistance 2 (Sr2), C6K1C2 and C6K2C1 for Lr34, and Lr46g22K3 for Lr46 were redeveloped, the genotyping results were clear; KASP markers Lr46_JF2-2 for Lr46, TM4 and TM10 for Lr67 were also converted into derived cleaved amplified polymorphic sequences (dCAPS) markers Lr46Rdcaps, TM4dcaps, and TM10dcaps, respectively. dCAPS markers would release the machinery requirement and lower the application threshold. In conclusion, this study enriches marker type and quantity, and provides convenience for marker assistant selection of pleiotropic disease resistance genes. It also has important reference value for KASP marker development and improvement.

Low temperature is one of the critical environmental stress affecting the growth and development of sorghum (Sorghum bicolor) in the cold region. In order to study the physiological changes of sorghum seedlings leaves and the expression of related genes under low temperature stress, the cold-resistance variety 'P61' and the cold-sensitive variety 'H21' were selected as the test materials, which were subjected to low-temperature treatment at 4 ℃ for 0, 1, 3, 5 and 7 d, respectively. The results showed that the chlorophyll fluorescence parameters (Fv/Fm) and relative chlorophyll content (SPAD) decreased significantly with the extension of the low-temperature treatment time, whereas the decrease was not obvious in the cold-tolerant variety 'P61'; the expression of genes related to photosynthesis were down-regulated with the prolonged stress time, and the expression level was higher in resistant varieties; the activity of superoxide dismutase (SOD) and peroxidase (POD) was elevated after 5~7 d treatment. At 7 d, the SOD activity of 'H21' decreased by 3.3%, the SOD activity of 'P61' increased by 15.2%, and the POD activity of 'H21' and 'P61' increased by 7.4% and 9%, respectively. The content of malondialdehyde (MDA) and proline (Pro) increased. The MDA content of 'P61' and 'H21' increased by 36.5% and 1.7%, respectively, the content of Pro increased by 69% and 230.5%. A set of 8 SbSODs genes were screened from transcriptome database, the number of SbSODs up-regulated at low temperature for 3 d were larger than those treated for 1 d, and the expression of SbSOD5 was up-regulated in the low-temperature treatment of both varieties for 1 d and 3 d; 7 sorghum PODs genes were screened, and the most of the genes (5 out of 7) were up-regulated in both varieties when treated for 1 d, Sobic.009G055100 was up-regulated in both varieties after 1 d and 3 d treatment; a total of 11 SbLOXs were screened, and a number of up-regulated SbLOXs (lipoxygenase) were observed at 1 d time point, and SbLOX9 was up-regulated in both varieties under 1 d and 3 d treatments; 2 sorghum P5CS genes were selected, of which Sobic.003G356000 was significantly up-regulated at different time points in two varieties, and a high expression level (Log₂FC>6 ) was detected in 'P61' when treated for 3 d. This study provides a reference for further research on the physiological regulation of plants under low temperature stress and the transcriptional regulation of related genes.

Plant calmodulin-binding transcription factor (CAMTA) plays important roles in hormone signal transduction, developmental regulation, and low-temperature stress. In this study, the genome-wide identification of CAMTA genes were carried out in tomato (Solanum lycopersicum), the pivotal information of the family members was comprehensively analyzed by bioinformatics technology, and qRT-PCR was used to analyse the expression pattern of them under low-temperature stress. The results showed that there were 18 CAMTA gene family members were identified in tomato genome, which were distributed on 9 chromosomes and existed in one pair of segmental duplication gene. Based on phylogenetic analysis, the SlCAMTA members were clustered into 6 groups. There are 49 cis-acting elements related to plant hormone, stress, light response and tissue-specific expression were identified in the promoter region. Protein-protein interaction network showed that some of SlCAMTA proteins were closely related to Arabidopsis reported proteins. Most of the SlCAMTAs were expressed in various tissues, and more than half of the CAMTA genes had molecular functions and constituted the basic components of cells. Especially, SlCAMTA10 participated in 11 plant biological processes. Under low-temperature stress, 14 SlCAMTAs were significantly up-regulated expression, and the expression level of SlCAMTA02 reached the highest at 12 h (396.53), which was 396 times as compared with the control. Accordingly, this study provides a reference for further excavation of the gene function in tomato CAMTA family and the response mechanism to low-temperature tolerance.

BABY BOOM (BBM) proteins are plant-specific transcription factors belonging to APETALA2/ethylene-responsive factor (AP2/ERF) family that may play an important regulatory role in somatic embryogenesis. By means of homology-based cloning, GhBBMa (GenBank No. MW836956) and GhBBMd (GenBank No. MW836957) were obtained from upland cotton (Gossypium hirsutum) cultivar YZ-1. The resequencing and phenotypic data of three natural populations, and the genome sequence of cultivar ZM24 were collected to characterize BBM genes and corresponding proteins. Especially, the expression profiles were analyzed by RNA-seq dataset and qRT-PCR assay. GhBBMa and GhBBMd were located on chromosomes A08 and D08. The opening reading frame (ORF) of GhBBMa/d were both composed of 9 exons, correspondingly with 2 028 bp and 2 025 bp in coding sequence (CDS), encoding 675 and 674 amino acids, respectively. The GhBBMa/d proteins contained two AP2 domains and excluded signal peptide or transmembrane domains. The bioinformatics subcellular location indicated that GhBBM appeared to accumulate in the cytoplasm. The phylogenetic tree showed that the GhBBMs were closely related to that from Theobroma cacao, which both belonged to the Malvales. The resequencing data of the natural populations showed that the sequence polymorphisms of GhBBMa and GhBBMd were 1.222 and 1.422 SNP/kb, respectively. The linkage disequilibrium (LD) decay distances were over 5.54 kb and 12.76 kb for GhBBMa and GhBBMd, respectively, indicating that GhBBMd was subjected to more evolutional selective pressure than GhBBMa. Association mapping showed that GhBBMa and GhBBMd had significant phenotypic effects on traits such as micronaire values and so on. RNA-seq dataset showed that the expression level of GhBBMa and GhBBMd genes was high in roots and ovules, but was very low in other tissues. GhBBMd was more vigorously transcripted in embryogenic callus than that in non-embryogenic callus. qRT-PCR assays showed that GhBBMa and GhBBMd had the highest expression accumulation in 30 d embryogenic callus and -3 days post anthesis (DPA) ovules. In summary, GhBBMa and GhBBMd, were cloned with low sequence diversity in DNA and substantive phenotypic effects on several traits. Two genes exhibited divergent expression abundance at different stages of somatic embryogenesis. According to the integrated results from RNA-seq dataset and qRT-PCR assay, GhBBMa/d might function in triggering embryogenesis. This study will facilitate the explore of mechanism of GhBBM involved in somatic embryogenesis.

Dehydration-responsive element binding protein (DREB) belongs to a subfamily of ethylene response factor (ERF) family. The cloning and functional analysis of DREB gene family is helpful for further revealing the regulation mechanism of APETALA2 (AP2)/ERF superfamily transcription factors in sunflower (Helianthus annuus) under biotic and abiotic stress. Based on the known sequence obtained from salt-induced differentially expressed genes in sunflower, the 3' and 5' ends of the sequence were amplificated and assembled into full cDNA sequence of 847 bp (GenBank No. MH085932). The cDNA contained an open reading frame of 468 bp which encoded 155 amino acids. The isoelectric point and molecular weight of the protein was predicted to be 5.21 and 37.5 kD, respectively. The amino acid sequence contained a conserved AP2/ERF DNA binding domain, including conserved YRG element and WLG motif, and the 14th and 19th amino acids in the AP2/ERF binding domain was valine and glutamate, respectively, implying that the gene belonged to DREB subfamily. Evolutionary analysis of transcription factors of AP2/ERF subfamily in Arabidopsis thaliana further indicated that the transcription factor belonged to A5 group of DREB subfamily．Blast analysis showed that the gene had a high similarity with many reported nucleotide sequences and deduced amino acid sequences of ERF family members in other plants. The full-length gDNA sequence of the gene was cloned by PCR method according to the full-length cDNA sequence of HaDREBA5 with the length of 468 bp from start codon to stop codon (GenBank No. MH085933), which had no introns. qRT-PCR study indicated that the expression of HaDREBA5 in sunflower could be induced by pathogen, mechanical damage, low temperature, NaCl and salicylic acid stress and showed different expression patterns under different types of stress. HaDREBA5 was expressed in roots, hypocotyls and leaves of sunflower and had organ expression specificity. The subcellular localization displayed that it was distributed in the nucleus of onion (Allium cepa) cells. The overexpression of HaDREBA5 in tobacco (Nicotiana tabacum) plants could enhance the tolerance to low temperature, drought and salt stress. Under NaCl stress, both the content of chlorophyll, soluble protein and proline and the activity of peroxidase (POD) and superoxide dismutase (SOD) increased; the expression of stress-related genes such as P5CS (Δ′-pyrroline-5-carboxylate synthetase), POD, MnSOD and GuZnSOD were also up-regulated in transgenic tobacco plants. The cloning and functional study of HaDREBA5 gene will help to understand the molecular mechanism of sunflower stress resistance, and provide gene resources and reference for stress resistance breeding in sunflower or other crops.

Mitochondrial membrane-binding proteins (MMBPs) encoded by nucleus genes involve in cellulose accumulation. However, more MMBPs need to be identified and characterized to disclose its role in cellulose accumulation metabolism. In this study, St536 (ID: PGSC0003DMG400012536), which was screened from the previous transcriptome analysis and encoding membrane-binding protein, was cloned in potato (Solanum tuberosum). Sequence analysis showed that the ORF of St536 gene was 648 bp, encoded 215 amino acid. Subcellular localization showed that the protein encoded by St536 gene was detected in the mitochondrial membrane. St536 overexpression vector was constructed and transformed into tobacco (Nicotiana tabacum) to obtain St536 overexpression lines of tobacco. Mophological assay and physiological analysis on the St536 overexpression lines showed that cellulose accumulation and the ratio of stomatal and guard cell relative vertical length (GL/SL) in the overexpression lines reduced 20% and 17%, respectively, comparing with wild type (WT). Stomatal conductance and photosynthetic rates in the overexpression lines were 2.4 and 2.5 times of the wild type, respectively. The result indicates that St536 overexpression decreased cellulose accumulation, lead to stomatal open and close easier, and enhanced stomatal conductance and photosynthesis rate. The study confirmed that St536 was involved in leaf cellulose accumulation, and provides basic data for further revealing the mechanism of cellulose accumulation.

BLM helicase (bloom helicase, BLM) plays an important role in DNA replication, repair, recombination, and maintenance of telomere stability. When the BLM helicase gene is mutated, it is prone to cause BLM syndrome and even cancers. To explore the expression patterns of long coding RNAs related to the BLM helicase gene. In this study, miR-27b-3p, which was related to the regulation of BLM genes, though the self-developed progame SWChen2.3 to screen the LncRNAs associated with the BLM genes, and an analyzed the BLM-associated with the LncRNAs in different prostate cancer cell lines PC3、Lncap、22RV-1 and normal fibroblst cell line WPMY-1, candidate LncRNAs were cloned, and their secondary structure and ORF were predicted, and target sites related to BLM gene were analyzed. NONHSAT203515.1 (m130127_152334_00126_c100) (named BLNC203) and NONHSAT246735.1 (lnc-ZBTB201:1) (named BLNC246) were selected and employed as research objects. The qRT-PCR method was used to detect the expression of BLNC203 and BLNC246 in 3 different prostate cancer cells and a normal epithelial cell line. The qRT-PCR results showed that the 2 LncRNAs were expressed in different prostate cancer cells, the expressions of BLNC203 in PC3 and 22Rv-1 were significantly higher than that in normal cells (P<0.01), while the differences of Lncap expression were not significant among these cells (P>0.05). The expressions of BLNC246 in PC3 and Lncap cells were significantly lower than that in normal cells (P<0.01), and while the differences of 22RV-1 expression were also not significant among these 22RV-1 cells (P>0.05). BLNC203 had 3 ORFs, BLNC246 had 1 ORF. the minimum free energy of the secondary structure of BLNC203 was -239.10 kcal/mol, and the minimum free energy of BLNC246 was -85.30 kcal/mol. The results of this study will provide a theoretical basis for further research on the mechanism of LncRNAs/BLM on the proliferation of prostate cancer cells.

The aim of this study was to clone the gene sequence of Aquaporin-1 (AQP1) in yak (Bos grunniens) and study the expression of AQP1 in yak testes of different ages. Testicular samples of newborn (3 day), young (1 year old), adult (4 years old) and old (8 years old) yak were collected. The full-length cDNA sequence of AQP1 gene was amplified by RT-PCR technology and analyzed by bioinformatics. qRT-PCR, Western blott and immunohistochemistry methods were used to detect and locate at the gene and protein level. The qRT-PCR results showed that as the yak grew older, the expression level of the yak AQP1 gene showed a trend of rising first and then falling, reaching the highest in the young testis; the expression level in the young group was significantly different from other groups (P<0.05), while the difference between the middle-aged group and the elderly group was not significant (P>0.05). Western blot results showed that as the yak grew older, the expression level of AQP1 protein also showed a trend of first rising and then falling, reaching the highest in the young testis; the expression difference between the young group and other groups was significant (P<0.05), while the differences between the newborn group, middle-aged group and old group were not significant (P>0.05). Immunohistochemical results showed that AQP1 protein is positively expressed in yak testes of different ages, and the expression sites are basically same, mainly expressed in sperm, sperm cells, spermatogenic cells, testicular stromal cells, supporting cells, and peritubular muscles like cells and seminiferous tubules in the basement membrane. The results of showed that the expression of AQP1 in the young yak group was significantly higher than that in the other groups (P<0.05). It is speculated that AQP1 plays an important role in the young yak testis. This study provides theoretical basis for further research on the mechanism of AQPs in the yak testis.

The wool yield of transgenic Chinese Merion fine wool sheep (Ovis aries) with insulin-like growth factor I (IGF-I) gene was significantly increased. In order to find out the change of skin protein expression related to wool production, data independent acquisition (DIA) proteome sequencing technology and bioinformatics analysis were used to analyze the total skin protein of transgenic sheep and wild-type sheep, which aimed to find specific proteins related to wool growth characteristics. Firstly, DIA proteomics method and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to sequence the tissues of IGF-I transgenic sheep and wild type sheep. The sequencing data were normalized, differentially expressed proteins were analyzed, Gene Ontology (GO) and KEGG enrichment analysis and candidate protein screening were carried out. The results showed that 2 914 proteins were obtained, including 930 differentially expressed proteins. Compared with wild type, 374 proteins were up-regulated and 556 proteins were down-regulated, and 18 differentially expressed proteins were related to wool traits. GO analysis showed that there were 24, 14 and 18 annotations to biological process, molecular function and cell component, respectively; KEGG pathway analysis showed that the differentially expression proteins were involved in 42 signaling pathways, including 8 signaling pathways such as serine/threonine kinase, secretory glycoprotein, gonadotropin releasing hormone, tumor suppressor protein, transforming growth factor-β, which was related to wool growth and development. This study is helpful to reveal the relationship between differential protein and wool traits and provides a reference for exploring the molecular mechanism of wool traits and molecular marker breeding.

Receptor tyrosine protein kinase 4 (ERBB4) and neuregulin 1 (NRG1) genes play an important role in the nervous system, the expression of ERBB4 gene in aggressive Cairna moschata was higher than that in normal group. In order to further explore the ERBB4 and NRG1 genes expression in nerve cells of Cairna moschata, pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 eukaryotic expression vectors were built and transfected to nerve cells of Cairna moschata, then the expression levels of ERBB4 and NRG1 genes and the downstream Src homolog and collagen homolog (Shc) protein and growth factor receptor bound protein 2 (Grb2) in the NRG1-ERBB4 signaling pathway were detected by qRT-PCR. The results showed that the expression of ERBB4 and NRG1 in the transfected pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 cells was significantly higher than those of pEGFP-C1 and pDsRED-C1 empty vector cells (P<0.01), indicating that pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 were successfully overexpressed in nerve cells of Cairna moschata; The expression of NRG1 in transfected pEGFP-C1-ERBB4 cells was significantly higher than that in transfected pEGFP-C1 empty vector cells (P<0.01), and the expression of ERBB4 in transfected pDsRED-C1-NRG1 cells was significantly higher than that in transfected pEGFP-C1 empty vector cells (P<0.01), it showed that the overexpression of ERBB4 and NRG1 both the were up-regulated by each other; Compared with the transfected pEGFP-C1 and pDsRED-C1 empty vector and the blank experimental group, the expression of Shc and Grb2 in transfected pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 cells were significantly up-regulated (P<0.01). The above results indicated that the expression up-regulated of ERBB4 or NRG1 significantly increased the expression of the other one, and the downstream genes Shc and Grb2 of NRG1-ERBB4 signaling pathway were increased in Cairna moschata nerve cells. The results of this study provide a basis for further exploring the mechanism of ERBB4 and NRG1 genes in the aggressive behavior of Cairna moschata.

Growth hormone-releasing hormone (GHRH) plays an important role in regulating the release of growth hormone in animals. In order to study the relationship between the GHRH gene polymorphism and growth traits in grass carp (Ctenopharyngodon idella), the cDNA and DNA sequence of GHRH gene (GenBank No. 2446387) were obtained by PCR amplification. Total length of cDNA and DNA sequence of GHRH were 602 and 5 244 bp, respectively, including 5 exons and 4 introns. Through direct sequencing, 12 SNP were identified in the DNA sequence, which located at 670, 1 798, 2 340, 2 782, 3 925, 4 227, 4 371, 4 420, 4 497, 4 976, 5 025 and 5 232 bp, respectively. After analysis with SNaPshot technique and General linear model, T+3925C, G+4227A, T+4420A, C+4497T, A+4976G and G+5025T were linked to form haplotype marker D1, and 4 SNPs were shown to have significant correlation with the growth traits in grass carp, which were C+1798T, T+2340C, A+2782T and haplotype D1 marker, respectively. The body weight in individuals with CC genotype in C+1798T was significantly higher than TT genotype (P<0.05); the body weight, body length, body height and head length in individuals with TT genotype in T+2340C were significantly higher than CT genotype (P<0.05); the body weight and body length in individuals with AA genotype in A+2782T were significantly higher than the other 2 genotypes (P<0.05); the body weight in individuals with EE genotype in haplotype D1 (TTGGTTCCAAGG) were significantly higher than the other 2 genotype (P<0.05). The linkage disequilibrium analysis of the 4 markers showed that the 2 combinations were in a strong linkage disequilibrium state (D'>0.8), which included 6 haplotypes: H1 (CTAE), H2 (TTTF), H3 CTTF), H4 (CTTE), H5 (CCTE) and H6 (TTAE). The body weight in haplotype combination of H1/H1 (CCTTAAEE) was significantly higher than that of other haplotype combinations (except H1/H3, P<0.01), which was 9.96% higher than the average body weight of the population. In conclusion, C+1798T, T+2340C, A+2782T, haplotype D1 markers and haplotype combination H1/H1 in GHRH were significantly correlated with growth traits, which provides candidate markers for marker assisted breeding in grass carp, and contributes to accelerate the breeding process of grass carp.

Head smut is a common and serious disease of corn. To clarify the genetic relationship of Sporisorium reilianum f. sp. zeae of different regions and different years in Shanxi Province, inter simple sequence repeats (ISSR) marker was used to analyse their genetic diversity, furthermore, Popgene Version 1.31, PowerMarker V 3.25, Mega 5.0 and Ntsys 2.10 softwares were used to estimate genetic diversity parameters, analyse genetic differentiation, and construct cluster maps. The results indicated that a total of 92 bands were detected using the 9 polymorphic primers screened, 90 (97.83%) among which were polymorphic, with an average of 10.0 polymorphic bands per primer. The average index of effective alleles (Ne), Nei's gene diversity (H), Shannon information (I) and polymorphism information content (PIC) were 1.360 0, 0.231 3, 0.370 7 and 0.443 7, respectively. All the tested strains were clustered into 9 ISSR groups at genetic similarity of 0.718. Strains from the same geographic origin were distributed into different IG groups, and strains from the same host variety in the same region were not preferentially clustered together. Comparing the population genetic diversity and variation of S. reilianum f. sp. zeae in 2015 and 2020, it was found that the genetic diversity parameters (PPB, Ne, H and I) showed a decreasing trend, and the genetic variation among geographic populations rose from 16.40% to 59.41%, while that within populations decreased from 83.60% to 40.59%. The gene flow changed from 2.549 1 to 0.341 6. All the results showed that rich genetic diversity existed among the tested S. reilianum f. sp. zeae strains. The ISSR groups of the strains had no correlation with the origin and host variety the population structure of S. reilianum f. sp. zeae changed significantly in 2020 comparing that in 2015. There are potential genetic differentiation factors among geographical populations and the stability of the populations has decreased significantly. The results provide an important theoretical basis for rational distribution of maize varieties and integrated control of head smut in Shanxi Province.

Wheat stem rot (WCR) is an important disease affecting wheat yield, Fusarium pseudograminearum is the main pathogen. The rapid and accurate identification of pathogens is significant for making reasonable control measures and resistant varieties breeding. In this study, elongation factor 1α (EF-1α) gene sequences of common Fusarium spp. were compared and specific primers F.pseudo-F3/R1 for F. pseudograminearum were designed, touchdown PCR were used to specifically amplify a 334 bp band from F. pseudograminearum, and no band amplified in F. graminearum, F. tricinctum, F. proliferatum, F. incarnatum and F. oxysporum. Wheat samples collected from Jining and Jiyang in Shandong province in 2019 and 2020 were detected using the site-specific PCR method which was just developed to test its validity. It was found that 28 fungi strains were isolated from infected samples from Jining in 2019, in which F. pseudograminearum was amplified with aimed band of 334 bp using primers F.pseudo-F3/R1, while no band from other fungi including F. graminearum, F. equiseti, Alternaria alternate, Bipolaris sorokiniana and Rhizoctonia cerealis. Samples were also collected from Jining in 2020 and DNA isolated directly from wheat stem with symptom of crown rot, then amplified using primers F.pseudo-F3/R1 and got a 334 bp aimed band of F. pseudograminearum, while other samples with symptom of root rot, leaf rust, powdery mildew or white-headed got no amplification band. The fungi isolated from stem of samples used above from Jining in 2020 were identified by sequencing analysis, and the result showed that F. pseudograminearum and F. graminearum existed in crown rot wheat sample, while F. graminearum and R.cerealis existed in other samples. The wheat stems of artificially and naturally infected by F. pseudograminearum from Jiyang in 2020 were amplified using primers F.pseudo-F3/R1 and got a 334 bp aimed band of F. pseudograminearum. The above results indicated that the specific PCR method could be used to detect F. pseudograminearum in stem directly, which provides a new method to rapidly identify F. pseudograminearum.

Hairy roots are a pathological phenomenon that occurs after plants are infected by Agrobacterium rhizogenes. It can produce the same or similar compounds as the original plant. Hairy root culture is an important method to produce plant secondary metabolites. In recent years, research on the strategy of using hairy roots to produce secondary metabolites has attracted more and more attention. This review mainly discusses hairy root culture conditions, expanded culture, and metabolic engineering. It introduces the research about plant secondary metabolism production based on the hairy root system, as well as the problems that need to be solved in the industrial production process. It is also explored that the future development direction of research on secondary metabolites produced by hairy roots. This article provides a reference for the hairy root production of secondary metabolites.

Genetically modified maize (Zea mays) CM8101 is one of the transgenic maize lines with commercial prospect developed independently in China. The research of quantitative detection method for CM8101 transgenic maize and its products is one of the technical requirements for the safety supervision of transgenic crops. In this study, real-time fluorescence quantitative primers and TaqMan probe were designed according to the 5' side sequence information of transgenic maize CM8101. After specificity, linearity, limit of detection, limit of quantification, accuracy, precision and reproducibility tests, a specific real-time fluorescence PCR quantitative detection method for CM8101 maize was established.The results of sample determination showed that this quantitative method had strong specificity and good accuracy, and could accurately quantify CM8101 transgenic components as low as 0.1%, and the detection limit could reach to 10 copies. In this study, an accurate quantitative detection method for CM8101 transgenic maize was developed based on qRT-PCR technology, which is expected to provide necessary technical support for CM8101 transgenic corn safety supervision.

Disease-resistant transgenic papaya (Carica papaya) has made great contribution in solving the problem of Papaya ring spot virus (PRSV). At present, the main transgenic papaya lines are '55-1', 'GM YK' and 'Huanong No.1'. In order to supervise the disease-resistant transgenic papaya products in the market and protect consumers' right to know and right to choose, it is necessary to establish more efficient and accurate detection method for transgenic papaya. This study analyzed the major genetically modified (GM) papaya in the world and established a simple and efficient multiplex PCR detection method for specific sequences of the papaya endogenous reference gene Papain, screening marker gene NPTⅡ, event-specific sequences of GM papaya lines '55-1', 'GM YK' and 'Huanong No.1'. The test results suggested that the established multiplex PCR detection method had high specificity and sensitivity. Samples could be judged whether they were transgenic papaya and which type of transgenic papaya were contained by only once PCR assay and electrophoresis. This study provides technical support for the composition detection and identity verification of transgenic papaya.

Bovine respiratory disease syndrome (BRDC) is a serious disease of cattle caused by a mixture or secondary infection of bacteria, viruses and environmental factors, which seriously endangers the healthy development of cattle industry. Based on the glycoprotein B (gB) gene of Infectious bovine rhinotracheitis virus (IBRV) in GenBank (GenBank No. NC0847.1), lipoprotein 48 (p48) gene of Mycoplasma bovis in GenBank (GenBank No. DQ020482.1) and Khe gene of Klebsiella pneumoniae in GenBank (GenBank No. Kx842080.1), three pairs of primers were designed to amplify the expected fragments of 727, 421 and 192 bp for Infectious bovine rhinotracheitis virus, Mycoplasma bovis and Klebsiella pneumoniae, respectively. The optimization of annealing temperature and primer concentration in PCR amplification, the specific test and clinical sample detection were carried out, and the multiplex PCR and sample evaluation were established. The results showed that the optimum annealing temperature was 52.7 ℃ in the range of 48~54 ℃, and the optimum primer concentration of IBRV, M. bovis , K. pneumoniae was 1, 1, and 15 mol/L in the range of 1~15 mol/L, and the lowest detection rate of IBRV was 10³ TCID₅₀, the lowest detectable amount of K. pneumoniae was 8.1×10¹ cfu and the lowest detection level of M. bovis was 6.5×10¹ cfu, and no cross reaction was found with Pasteurella multocida, Salmonella typhimurium, Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Serratia marcescens, Pseudomonas aeruginosa and Bovine diarrhea virus. The results of clinical samples were consistent with the isolation and identification of pathogens. The establishment of multiplex PCR method provides strong technical support for the diagnosis and control of bovine respiratory disease syndrome caused by these three pathogens.