In this study,the information that contained phylogenetic relationship and distribution of apple potassium(K+)transporter genes was analyzed based on Malus genome database,and their expression characteristics were explored by means of qRT-PCRs in apple. The results showed that 65 K+ transporter genes were identified,and they were divided into 4 families:CHX family,HAK family,HKT family and KEA family,including 33,24,1 and 7 members,respectively. These genes are highly homologous with Arabidopsis thaliana K+ transporter genes. Chromosomal localization exhibited that these K+ transporter genes were distributed in the thirteen of seventeen apple chromosomes unevenly. Quantitative expression analysis showed that most of the K+ transporter genes had differential expression patterns. The thirty-two genes had higher expression abundance in root while 12 genes in mature leaves,11 genes in shoot tips and 19 genes in fruit. The results lay a basis for revealing the function of K+ transporter genes in apple.
A gene named MdDRB1 cloned from Malus × domestica‘Royal Gala’was analyzed. To understand sequence characteristics of the gene,prokaryotic expression and subcellular localization were carried out. In addition,real-time quantitative PCR were performed to determine the expression levels of MdDRB1 in response to various abiotic stresses. Finally,MdDRB1 was genetically transformed into apple seedlings to identify its function. The analysis of gene structure revealed that there were two introns and three exons in genomic sequence of MdDRB1. Subcellular localization showed that MdDRB1 were mainly distributed in the nucleus,a small number distributed in the cytoplasm. Then,the induced protein showed that MdDRB1 fusion protein existed in the form of inclusion body. Furthermore,the miRNA related to resistance expression level raised in antisense transgenic apple callus. In addition,real-time quantitative PCR found that the MdDRB1 gene expression level significantly raised after treated with PEG,NaCl,ABA and 4 ℃. Overexpression of MdDRB1 remarkably increased the tolerance of transgenic apple seedlings to abiotic stresses. These results indicated that MdDRB1 might be involved in the response of apple to abiotic stresses.
The effects of high temperatures on leaf traits,chloroplast ultra-structures,gas exchange,chlorophyll fluorescence parameters,antioxidant enzyme activity,pigment contents,and electrolyte leakage of three north highbush blueberry cultivars(‘Bluecrop’,‘Duke’and‘Brigitta’)were examined with four growth chambers by controlling growth temperatures at 25,30,35 and
CitMYB22,an member of MYB family gene,was cloned using RT-PCR from leaf of Citrus junos Sieb. ex‘Ziyang’. Comparison between cDNA and genomic DNA sequences showed that two introns were existed in CitMYB22 gene. The 696 bp length ORF of CitMYB22,encoding a putative protein of 231 amino acids. The conserved R2R3 domain was observed in the amino acid structure sequence of CitMYB22. Phylogenetic analysis revealed that CitMYB22 shared the highest amino acid identities with homologous Arabidopsis R2R3-MYB transcription factor AtMYB15. A 2 500 bp long promoter of the CitMYB22 gene was isolated,sequence analysis showed that the promoter harbors multiple stress- responsive cis-elements,such as HSE,SARE and MYS. The green fluorescent protein transient expression assay revealed that the CitMYB22 protein was localized in nucleus location. qRT-PCR analysis indicated that the higher expression of CitMYB22 gene was in leaves and flowers than in roots and fruitlets. Moreover,CitMYB22 was induced by exogenous hormone abscisic acid(ABA),salicylic acid(SA),methyl jasmonate (MeJA),1-amino cyclopropane carboxylate(ACC)and physiological stresses of salinity,dehydration and low temperature(
The cross-compatibility experiments of 91 inter-specific combinations and 21 inter-generic combinations were carried out among 7 Eriobotrya plants and 2 related genera(Raphiolepis indica Lindl. and Photinia serrulata Lindl.)by emasculation,bagging,and artificial pollination. The results showed
To analyze genetic linkage and map the location of purple gene(BrPur)controlling purple inner leaves trait of Chinese cabbage,a F2 population constructed by the single-plant-selfing of F1 hybridized from homozygous purple heading line‘14S
In the present work,a WRKY transcription factor,termed BrWRKY75 was isolated from Chinese flowering cabbage. Analysis of deduced amino acid sequence and phylogenetic tree found that BrWRKY75 showed high homology with Brassica oleracea WRKY75 and Arabidopsis thaliana WRKY75,and belong to Ⅱ c sub-group. Real-time quantitative PCR displayed that BrWRKY75 was up-regulated during leaf senescence of Chinese flowering cabbage. Sub-cellular localization and transcriptional activity analysis revealed that BrWRKY75 is a nuclear protein with transcriptional repression activity. In addition,electrophoretic mobility shift assay(EMSA)confirmed that BrWRKY75 directly bound to the W-box(TTGAC)cis-element. Taken together,these results provide basis for the further investigation of the transcriptional regulation mechanism of Chinese flowering cabbage leaf senescence.
Basis on the RNA-seq analysis,the relative expression and function of carrot SUPPRESSOR OF OVEREXPRESSION OF CONSTANTS 1(SOC1)homologues under photoperiod treatment were investigated with wild sensitive species‘Songzi’and cultivar‘Amsterdam’which was tolerant to premature bolting. There were two carrot SOC1 homologues:DcSOC1-1 and DcSOC1-2,which were annotated with complete open reading frames(ORF)encoding 217 and 211 amino acids respectively,which both containing MADS-box and K-box domain and a conserved SOC1 motif at the carboxy terminus. Phylogenetic analysis showed that DcSOC1-1 and DcSOC1-2 were closely related with kiwifruit AcSOC
This research aims at identifying the currently prevailing viruses infecting Hami melon in Xinjiang and their genetic variations for providing scientific basis to prevent these virus diseases. Field survey was conducted in Turpan,Shanshan and Changji and samples with viral symptoms were collected for the lab tests. Both DAS-ELISA and RT-PCR were applied for the detection and molecular identification of 7 common viruses as well as Cucumber green mottle mosaic virus(CGMMV)the important quarantine virus infecting Hami melon. The results showed that the detection rates of Cucumber mosaic virus(CMV),Watermelon mosaic virus(WMV),Zucchini yellow mosaic virus(ZYMV),and Melon necrotic spot virus(MNSV)amongst the 121 detected samples were 70.2%,62.0%,37.2%,and 2.5% respectively,whereas Squash mosaic virus (SqMV),Papaya ring spot virus(PRSV),CGMMV and Tobacco necrosis virus(TNV)had not detected as positive. CMV,WMV and ZYMV are widely distributed in the field. Mixed infection with two or three different viruses on the same individual host was common with an occurrence of 60.19%. Based on the CP gene amplification,sequencing and phylogenetic analysis,CMV isolates from Hami melon in Xinjiang were categorized into subgroupⅠB. Although WMV,ZYMV and MNSV had high homology with other reported strains in CP gene nucleotide sequence,there were still some variations. ZYMV and WMV had a significant geographical differentiation.
Based on the transcriptome data of Colocasia esculenta(L.)Schott,full-length cDNA of ApS1(ADP-glucose pyrophosphorylase small subunit 1)gene was cloned from‘Jingjiang Xiangsha Taro’,which consisted of 1 596 bp and encoded a protein of 532 amino acids,and tentatively designated as CeApS1(NCBI:Colocasia KU288757,unreleased). CeApS1 contained a NTP_transf_3 domain(Gly104–Pro306)and a PbH1 domain(Gly473–Ser515). This gene encoded a small subunit of ADP-glucose pyrophosphorylase,and isoelectric point and molecular weight were 6.73 and 57.6 kD,respectively. Results displayed ApS1 existed in monocotyledons and dicotyledons. BlastX showed that ApS
In this experiment,osmotic regulation substances,antioxidant substances and ion were compared between Lilium regale Wilson(salt tolerance)and‘Sorbonne’(salt sensitiveness). The results showed that the change trend of these substances was the same in two lilies and L. regale
The two years old Fuji/Malus robusta biennial plants were selected as test materials,and potted in sand fertilized with Hoagland’s solution. The solution included five different Ni levels. After 20 days of spraying 15N-labeled urea into apple leaves,effects of Ni on absorption,transformation,and distribution of urea in the apple trees were detected,meanwhile effects of different Ni levels on nickel content,glutamine synthetase(GS)activity and superoxide dismutase(SOD)activity of leaves were analysed in the study. The results showed that,with the increase of nickel levels,the nickel content in apple leaves improved correspondingly. Under low level of nickel treatment(0.5,1.0,2.0 mg · plant-1 NiSO4 · 6H2O),the GS activity and SOD activity in leaves were higher than those of the control,and the treatment with 1.0 mg/plant NiSO4 · 6H2O had the best effect in the study. However,the GS activity and SOD activity in leaves under high level of nickel treatment(10 mg · plant-1 NiSO4 · 6H2O)were significantly lower than those of the control. The 15N-urea utilization ratio of 0.5,1.0 or 2.0 mg · plant-1 NiSO4 · 6H2O treatment was significantly higher than that of the control,and the 15N use efficiency of 1.0 mg NiSO4 · 6H2O treatment was up to 48.74%,which was 1.71 times as many as that of the control,but the 15N use efficiency of 10 mg · plant-1 NiSO4 · 6H2O treatment reduced by 38.26% compared with the control. Nickel also had obvious effect on 15N distribution ratios in apple trees. The 15N distribution ratio in roots was the highest,followed by leaves and stems in terms of 1.0 mg NiSO4 · 6H2O treatment. However,15N distribution to leaves was the highest of the other treatments. Overall,the results suggested that appropriate nickel level could increase the GS and SOD activities,retard senescence of apple leaves,and improve the absorption and utilization of urea,but high nickel level could result in the opposite effect on utilization of urea.
Simple sequence repeat(SSR)markers were employed to investigate the genetic diversity and genetic structure of 280 individuals sampled from 14 natural populations of Prunus pseudocerasus. A total of 80 alleles of 11 loci were detected,and the number of alleles per locus ranged from 4 to 13. The relatively high levels of gene diversity(h,0.5431–0.7151)and Shannon’s diversity(I,0.9057–1.4684) revealed relatively rich genetic diversity among the 14 Prunus pseudocerasus populations. The hierarchical analysis of molecular variance(AMOVA)revealed the genetic differentiation mainly within populations(70.00%). Mantel test revealed a significant correlation between geographic and genetic distances(r = 0.472,P = 0.011). Therefore,the genetic diversity was high both at the population level(PPL = 100%,h = 0.643,I = 1.207)and the species level(PPL = 100%,h = 0.743,I = 1.591).
A pot experiment was conducted to determine the impacts of the low,middle and high concentrations of 4-hydroxybenzoic acid on the chlorophyll content,photosynthesis gas exchange parameters and chlorophyll fluorescence parameters of the different strawberry(Fragaria × ananassa Duch.) cultivars(‘Tutela’,‘Toyonoka’and‘All Star’). The results showed that the content of chlorophyll a,chlorophyll b and carotenoids of three strawberry cultivars were reduced under the 4-hydroxybenzoic acid stress with the largest decline of‘All Star’. The net photosynthetic rate and transpiration rate of‘Toyonoka’with a little increase under the lowest concentration of 4-hydroxybenzoic acid,while the net photosynthetic rate,stomatal conductance and transpiration rate of‘Toyonoka’,‘Tutela’and‘All Star’Were all decreased under the 4-hydroxybenzoic acid stress,and the greater decline the higher concentration. The initial fluorescence(Fo)and non-photochemical quenching coefficient(qN)were significantly increased under 100 mg · kg-1 and 200 mg · kg-1 4-hydroxybenzoic acid stress. Under the highest concentration of 4-hydroxybenzoic acid stress,the initial fluorescences(Fo)of‘All Star’,‘Toyonoka’and‘Tutela’were increased by 15.1%,21.5% and 9.3%,respectively,and non-photochemical quenching coefficient of the three strawberry cultivars were increased by 131.3%,190.5% and 26.6%,respectively. The maximum fluorescence(Fm),maximal photochemical efficiency(Fv/Fm),efficiency of excitation capture of open PSⅡ center(Fv′/Fm′)and effective quantum efficiency of PSⅡ(ΦPSII)of three strawberry cultivars were firstly increased with the increasing concentration of 4-hydroxybenzoic acid and then decreased. The comprehensive advantage of chlorophyll fluorescence parameters of three strawberry cultivars was‘Tutela’>‘Toyonoka’>‘All Star’. Low 4-hydroxybenzoic acid concentration(50 mg · kg-1) didn’t affect significantly the dry matter accumulation of‘Tutela’and‘Toyonoka’,but the aboveground and underground dry quality,and ratio of root/shoot per plant of three strawberry cultivars were significantly decreased in other treatments. The photosynthetic pigments,photosynthesis and chlorophyll fluorescence characteristics of‘Tutela’were less influenced by 4-hydroxybenzoic acid stress than‘All Star’and‘Toyonoka’. So a higher photosynthetic performance might be an important physiological trait of resistant cultivars in the adaptability to replant disease.
Pollen grains of 8 persimmon species composed of 7 known species and 1 unknown resource were collected from National Field GenBank of China. Pollen grains were examined using scanning electron microscopy,to study pollen morphology and taxonomic status. The results showed that:(1)Pollen morphology observation revealed the grains pretreated in Diospyros have three germinal furrow and the shape of grains are nearly spherical. The grain is round in polar view. Pollen size was measured by length of polar axis and equatorial axis. The value of pollen size was between 24.7 µm and 52.8 µm in polar axis and between 23.8 µm and 47.9 µm in equatorial axis. In germinal furrow,there are many granular spiral units. Exine of pollen are striped ornamentation like grain. The width of stripe and length of germinal furrow among these species exist significant differences. In addition,surface of pollen have obvious or not obvious perforations.(2)D. lotus L.,D. glaucifolia Metc.,D. oleifera Cheng. belonged to groupⅠ;Nishimurawase,D. virginiana,male strain No. 2 formed group Ⅱ;Deyangshi performed the group Ⅲ. The classification using clustering analysis is positively correlated with their ploidy levels. (3)Pollen morphology of Deyangshi is significantly different from other materials,and the results laid the foundation of palynology for taxonomic status of Deyangshi.
The main sugars and organic acid components in fruits of 20 Chinese jujube varieties of different development stages were determined to compare the difference among varieties and reveal the dynamic changes. The result indicated that major components of sugars in Chinese jujube fruits were sucrose,fructose and glucose,and the sucrose content was the highest,followed by fructose and glucose,it was sucrose accumulation type. Total sugar and each component contents had significant differences in different cultivars’ fruits.‘Xinzheng Huizao’had the highest content of total sugar. With the development of the fruits,fructose,glucose and sucrose contents were accumulated constantly. At the early stage of the fruit development,fructose and glucose were main;while sucrose was main after white period. The major components of organic acid in Chinese jujube fruits were malic acid,quinic acid and succinic acid,and the malic acid content was the highest,so the malic acid was main. Total acid and each components content were significantly different among varieties. The total acid content of‘Binxian Jinzao’was the highest. Total acid,malic acid,quinic acid and tartaric acid contents were going up slightly along with the development of fruits,and citric acid content had no significant difference during different development stages. Succinic acid content was low in white period,which had extremely significant difference with crisp ripe and full ripeness. The sugars and organic acids correlation analysis showed that the total acid and malic acid,quinic acid;total sugar and sucrose,fructose,glucose;malic acid and total sugar,sucrose had very significant positive correlation. The citric acid and fructose sugar,glucose had significant negative correlation. The total acid and fructose,tartaric acid and ursolic acid had significant positive correlation.
The copy number of exogenous gene is an important factor influencing the expression and genetic stability of the target gene. Currently,transgene copy numbers by Southern blot,which is laborious and time-consuming,requires relatively large amounts of plant materials and more experimental technology. Here,based on quantitative real-time PCR(qRT-PCR)technology,a protocol for estimating transgene copy number in GM citrus was constructed. The results showed that the low copy number of exogenous gene in transgenic line was one,and the highest copy number was five. One,two and ≥ three copies detected accounted for 40.0%,18.0%,42.0%,respectively,of the total. The exogenous gene copy in the transgenic lines was further confirmed by Southern blot,and the copy number was consistent with the above two methods,although the copy number by qRT-PCR analysis in a few of plants was more than that by Southern blot. Therefore,the evaluation result with Real-time fluorescent quantitative PCR method was more accurate and reliable in comparison to that with Southern blot. And transgenic lines with a copy number of 1–2 could be used as an experimental material for the evaluation of the GMO bio-safety.
A promising biocontrol agent HL5-4,a strain with strong effect on tomato gray mold,which selected from the rhizosphere of healthy tea plant. In vitro and in vivo experiments were used to determine its antifungal effect and colonization ability. The results showed that,HL5-4 inhibited mycelial growth and conidia germination of Botrytis cinerea in vitro experiment,the inhibition zone of the mycelial growth was
In order to develop a reverse transcription loop-mediated isothermal amplification(RT-LAMP)assay for rapid and sensitive detection of Cucumber mosaic virus(CMV)from different kind of vegetables,four primers were designed according to the coat protein(CP)gene of CMV. The reaction conditions of RT-LAMP,including the concentrations of dNTPs,Mg2+,reaction temperature and reaction time were optimized. The specificity and sensitivity of RT-LAMP were testified. The optimum conditions for RT-LAMP can carried out under the concentrations of 1.2 mmol · L-1 dNTPs and 6 mmol · L-1 Mg2+. RT-LAMP reaction was carried out at
‘Hanlü’is a new Chinese cabbage hybrid which is developed by crossing a self-incompatible line‘WC-
‘Xiyuan
‘Jinhoutou
Antirrhinum‘Xingyue’is derived from the cross of female‘Yellow General’and male‘Pink General’. It belongs to a standard cut flower cultivar. The plants are 105–
‘Mengxing’is a new Michelia cultivar which was selected from hybrid crossing Michelia chapensis Dandy× M. crassipes Law. It is an evergreen arbor with oblong obovate and thiny leathery leaves whose apexes are acuminate. Inner tepals are with deeper purplish red mottling compared with outer tepals. Androecium is also purplish red. The florescence can start from March until April. The cultivar which likes the warm and moist climate can be widely planted in the south of the
‘Guangfangfeng