The cytokinin O-glycosyltransferase gene MdZOG1 was cloned by combination of the homologous clone and PCR technology from apple(Malus × domestica‘Gala’). The Open Reading Frame (ORF)of MdZOG1 contained 762 bp,encoding a protein of 253 amino acid residues. Phylogenetic analysis showed that MdZOG1 had the highest evolutionary relationship with PbZOG1. The transcriptional analysis results showed that expression of MdZOG1 was mainly in root and stem,the expression of MdZOG1 in flower and fruit tissues was lower than other tissues. Transgenic Arabidopsis and tobacco were generated by Agrobacterium-mediated genetic transformation. Ectopic expression MdZOG1 in Arabidopsis and tobacco significantly enhanced resistance to water stress,indicating that cytokinin O-glycosyltransferase played an important role in plant drought stress.
To explore physiological and molecular mechanisms of apple dwarfing rootstock,we analyzed the characteristics of physiochemistry,including membrane injury,reactive oxygen metabolism and osmotic adjustment substances accumulation,and transcriptome profiling occurred in‘71-3-150’,a cold-resistant apple dwarfing rootstock originated from Russia,exposed to cold stress(4 ℃)for 0,2,6,12 and 24 h. We found the immediate antioxidant and osmotic adjustment response of the rootstock to cold stress emerged during the first 2 h of cold exposure. Activities of reactive oxygen-scavenging enzymes SOD and CAT,and contents of main osmotic adjustment substances plateaued after 6 h of cold. Meanwhile,aggravating trend of membrane injury was mitigating,indicating the rapid physiological response of the rootstock to cold. The initial 6 h could be critical period for cold acclimation of the rootstock. Based on transcriptome analysis,491,1 115,828 and 1 047 up regulated differentially expressed genes(DEGs)and 537,688,708 and 1 016 down regulated DEGs were identified in 2,6,12 and 24 h samples respectively,compared with 0 h sample. The common 115 up and 136 down regulated DEGs showed 6 expression patterns. Twenty-eight functional and regulatory DEGs were screened out. These genes involved in stress signals reception and transduction showed obvious up-regulated expression in early cold stress(0–2 h),while those that took part in osmotic adjustment,reactive oxygen metabolism,and membrane and protein protection expressed with various patterns. These results suggested the rapid and multiple molecular mechanisms of perception,transduction and responses to cold stress in cold acclimation of‘71-3-150’.
This experiment was conducted to explore the effect of carbendazim and bio-organic fertilizer on the apple replant disease. The Malus hupehensis Rehd. seedlings were planted in pots with apple replant soil. The effect of carbendazim combined with bio-organic fertilizer on Malus hupehensis Rehd. seedlings growth,and soil enviroment were studied. Compared with replant soil control,carbendazim,bio-organic fertilizer combined ap plication could improve root respiration rate,promote the growth of root system,improve the biomass of seedlings. The plant height and root respiration rate were increased 38% and 36% respectively. The bacterial/fungal ratios reached 19.07 times of control group of apple replanted soil,so that it became the maximum number among the three groups. The soil fungi community structure was changed,the combined application treatment had higher simpson index and lower shannon index,pielou index and margalef index;bio-organic fertilizer,carbendazim and bio-organic fertilizer combined application had significant effects on the soil enzymes. Therefore,carbendazim and bio-organic fertilizer combined application had the better effect on apple replant disease remission.
To investigate the effect of exogenous melatonin(100 nmol · L-1)on photosynthetic characteristics of grape seedlings under ozone stress(110 nL · L-1),potted Cabernet Sauvigon grapevine were used as materials to be irrigated with melatonin,gas exchange parameters and OJIP fluorescence transient were tested,and combined with chlorophyll fluorescence quenching analysis. The results showed that,the exogenous melatonin increased the content of chlorophyll a,chlorophyll b,carotenoids and the ratio of chlorophyll a/b under ozone stress,increased the net photosynthetic rate(Pn)of grape leaves and number of active reaction centers per unit area(RC/CSm),promoted the probability of a trapped exciton moves an electron into the electron transport chain beyond QA(Ψo),which reduced the photoinhibition under ozone stress. These results suggested that exogenous melatonin promoted the chlorophyll content under the ozone stress,reduced the damage of ozone stress on grape leaf photosynthesis by improving the energy distribution of grape leaf photosystem.
Pseudomonas fluorescensFD6 was isolated from the canola rhizosphere in Fujian. To define the inhibit ability of the strain FD6 to Monilinia fructicola,the causal agent of peach brown rot was detected using microscope concave slides method and on detached peach fruits. The pyrrolnitrin biosynthetic gene cluster was cloned using PCR amplification,and a prnA-deletion mutant was generated by homologous recombinant to identify its function. The results showed that strain FD6 could control completely brown rot on peach fruits. The bacterial culture of strain FD6 inhibited the spores of M. fructicola germination significantly by 93.18%,whereas the cultural filtrate of the strain FD6 only showed a lower inhibition to M. fructicola by 69.36%. The bacterium culture showed that high nematicidal activity to Meloidogyne incognita. The obtained 5 868 bp prn contained four genes prnA,prnB,prnC and prnD analysis indicated that prn of strain FD6 identified with that of P. protegens CHA0 and Pf-5 by 94%. prnA was required for PRN synthesis in P. fluorescens FD6,and involved in the biosynthesis of 2,4-diacetylphloroglucinol and pyoluteorin as well. organized in a single transcript unit. The phylogenetic
To analyze the photosynthetic physiological mechanism that exogenous salicylic acid(SA)relieves harm of tomato seedlings under the salt stress,‘Qinfeng Baoguan’tomato seedlings were taken as the testing material. Under hydroponic conditions,the chlorophyll fluorescence dynamics technology was used to research the effects of nutrient solution(1/4 Hoagland)applied with SA(200 μmol · L-1)on PSⅡ photochemistry activities of tomato seedlings and excitation energy distribution and antenna pigments absorb light energy utilization under stress of 100 mmol · L-1 NaCl. The results indicate that under the stress of NaCl in 15 d,there are PSⅡpotential photochemical activity(Fv/Fo),maximal photochemistry efficiency(Fv/Fm),antenna conversion efficiency(Fv′/Fm′),practical photochemical efficiency(ΦPSⅡ),photochemical fluorescence quenching coefficient(qP),quotient of absorbed luminous energy used for photochemical action(P)and chlorophyll fluorescence decay rate(Rfd)of seedling leaves applied with SA which increase in different degree. While there are also PSⅡnon-photochemical fluorescence quenching coefficient(NPQ),excitation pressure(1–qP),quotient of absorbed luminous energy used for antenna heat dissipation(D),photosynthetic function relative limit value(LPFD)and unbalanced distribution of coefficient deviation on excitation energy between two photosystems(β/α–1)which decrease evidently. But there is no obvious regularity on quotient of absorbed luminous energy used for non-photochemical dissipation(Ex)in reaction center of PSⅡ. These results above show that exogenous SA can decrease photoinhibition caused by salt stress through increasing PSⅡ photochemical activity,and then promote the growth and development of tomato seedlings.
Trehalose and the precursor trehalose-6-phosphate participate in responding to abiotic stress as the osmotic regulator and the signaling molecule. Trehalose-6-phosphate synthase(TPS)is one of the most important enzyme among the synthesis mechanism of trehalose-6-phosphate,and plays an important role in the metabolic pathway of trehalose. The over expression of TPS gene can improve crop’s tolerance to abiotic stress. Based on pepper genome database,11 genes of CaTPS family were identified by bioinformatics methods. Besides CaTPS3 contains one TPS and two TPP domains,the rest of 10 CaTPS family genes contain one TPS and one TPP domain. All 11 CaTPS family genes were classified into two classes according to their gene structure and phenogenesis. CaTPS1,CaTPS2 and CaTPS3 are grouped to ClassⅠ. CaTPS1 and CaTPS2 respectively contains 16 introns and 5 motifs from motif 1 to motif 5,while CaTPS3 contains 13 introns,one motif 3 and two motif 4. The rest eight genes from CaTPS4 to CaTPS11
SHORT VEGETATIVE PHASE(SVP)was a key regulatory factor in flowering-time control of Brassica juncea. In order to clarify the expression characteristics of SVP gene and the mediation sites of protein interactions between SVP with FLC in flowering pathways,we cloned SVP gene in‘Qingyejie’germplasm of Brassica juncea. Expression analysis of qRT-PCR revealed that SVP gene expressed in leaves as well as shoot apexes in the flowering pathways of vernalization and long-day photoperiod. SVP gene expressed at very low level(0.56 in stem tip and 0.35 in leaf blade) at early stage of vegetative phase. However,it significantly increased at early stage of the reproductive phase(The relative expression values in the vernalization pathway are 0.60 in stem tip and 1.27 in leaf blade,respectively. However,the values in the photoperiod pathway are 0.49 in stem tip and 1.42 in leaf blade). In stem tips,the expression of SVP was more sensitive to vernalization than that of long-day photoperiod. Contrarily,the sensitivity of SVP gene expression in leaves was exactly reversed. Yeast two-hybrid experiments and β-galactosidase activity assays showed that FLC2 retained protein interactions withI-domain mutantof SVPE90L as well as K-domain mutants of SVPK104C and SVPH106I. However,the interacting strength of SVP/FLC2 was weakened by each of the three mutation sites above mentioned. Another K-domain mutant of SVPR137L failed to interact with FLC2 but still interacted with FLC1,FLC3,FLC4 and FLC5. It suggested that the interaction of SVP/FLC2 was specifically regulated by the 137th amino acid site of SVP. Sequence comparison showed that two different genes of FLC4 and FLC5 encoded the same FLC protein,which had only one variable amino acid site compared with FLC3. However,there are respectively 28,19 and 18 variable amino acid sites in FLC1,FLC3 and FLC4-5 compared with FLC2. Additionally,FLC2 had 11 unique amino acid sites which were different from any of other FLC family members such as FLC1,FLC3,FLC4 or FLC5. Hence,we speculated that the variable sites between FLC2 and other FLC members probably contributed to the specific regulation of SVPR137L/FLC2.in B. juncea
Laboratory experiment and pot experiment were conducted to investigate effects of cinnamic acid treatments on seed germination,seedling growth,mycelia growth of Fusarium oxysporum f. fabae,occurrence of faba bean Fusarium wilt,rhizosphere microbial activity,diversity and community of faba bean. The results showed that seed germination and seedling growth were inhibited by cinnamic acid treatments,the higher the concentration,the stronger the inhibitory impacts. The most remarkable inhibitory impacts were observed on leaf number and root weight by cinnamic acid treatments. Mycelia growths of Fusarium oxysporum f. fabae were significantly promoted by cinnamic acid at low concentration and inhibited at high concentration. Disease index of faba bean fusarium wilt,amount of Fusarium oxysporum significantly increased and microbial activity(AWCD value),Shannon diversity index and substrate richness decreased with 100 and 200 mg · L-1 cinnamic acid treatments. Principal component analysis demonstrated that cinnamic acid treatment changed the rhizospheric microbial community composition obviously,and the carbohydrates,carboxylic acids,and amino acids are sensitive carbon sources for differentiating the changes of microbial community induced by cinnamic acid. Cinnamic acid reduced rhizosphere microbial diversity,promoted the pathogen infection of faba bean and increased the infection of Fusarium wilt of faba bean. Cinnamic acid is an autotoxin in the continuous cropping obstacle of faba bean.
Cucumis metuliferus is an important breeding resource for resistance to root-knot nematodes in Cucurbitaceae family. Peroxidase gene(CMPOD)was cloned from the CM3 line using primers of homologous sequences. There was 97% nucleotide sequence similarity in POD gene between CM3 and C. sativus‘9930’. POD gene mainly expressed in root of CM3 infected by Meloidogyne incognita,and the expression increased at the early stage of nematodes inoculation. POD gene expression reached the highest point at 3 d post inoculation. Moreover,temperature affected the POD gene expression which peaked at 20 ℃. When the temperature was above 20 ℃,POD gene expression reduced but the number of egg masses and galls increased with the rise of temperature. The trend showed a negative correlation between POD gene expression and the number of egg masses and galls. However,the nematode-resistance of CM3 was not compromised with increasing temperature and CM3 still showed resistance at 35 ℃. These results revealed that the root-knot nematodes resistance of CM3 was heat-stable,which could be utilized for crop breeding. Together,our studies provided an important basis to demonstrate the molecular mechanism of nematode-resistance of CM3.
In order to study the seedcoat characters and discuss their taxonomic significance,23 species from sect. Racemosae were sampled and investigated under SEM(Scanning Electron Microscope)in this study. The observation results indicated that the seeds of sect. Racemosae were ellipsoid,obovate,and oblong-ovate. The ratios of length to width are mostly between 1.4–2.3. Two types of micromorphogical character of seed surface were recognized:reticulate ornamentation and digitiform protrusive,and the later could be loosely or tightly digitiform protrusive. The seedcoat micromorphology characters of sect. Racemosae showed limited of correlation with the gross morphological characters. However,seedcoat micromorphological characters,such as granules present or not in the lumina,granules or pits emerge or not on the protrusive,the density of digitiform protrusive,and so on,are consensus within species and have taxonomical value in circumscription of species within sect. Racemosae.
The types,distribution of trichomes on the leaf epidermis and the change of external morphorlogy during secreting process of the glandular trichomes on the leaf epidermis of Elsholtzia stauntonii,and thefluorescence intensity change during the process of secreting flavonoids Benth. were investigatedwere observed by means ofoptical microscopy,scanning electron microscope and fluorescence nucriscipe with Neu’s reagent staining. The results showed that:(1)E. stauntonii possesses glandular trichome and non-glandular trichome on the leaf epidermis and glandular trichome includes capitate and peltate glandular ones. ,(68.14 ± 0.84)μm in diameter,while a matured capitate one had a smaller and hemispherical-shaped head with a diameter of(17.57 ± 0.48)μm. Non-glandular trichome includes mastoid,ramose and tubular non- glandular trichome.(2)Trichomes were distributed both on the abaxial and adaxial leaf epidermis of E. stauntonii,the abaxial epidermis was with more glandular trichomes than the adaxial one,and the peltate glandular trichomes were mainly distributed on the abaxial surface and the capitates were distributed in both surfaces of the leaves. While non-glandular trichomes were distributed on both leaf surfaces and more were found around the veins,along the leaf margin and at the base of blade.(3)Secretions from the glandular trichomes stored in the space between cytoderm of secretory cells and head cuticle. The pressure to the head cuticle increased with the secretions accumulating,which led to the head cuticle depressed or ruptured and the secretions released,at last the glandular trichomes broke down.(4)Both peltate and capitate glandular trichomes can secrete flavonoids. With the glandular trichome development and maturation,the flavonoids were accumulated gradually in the head,and then released to the external space.A maturing peltate glandular trichome had a larger and shield-shaped head
Simple sequence repeat(SSR)loci were searched and analyzed transcriptomic data of Prunus pseudocerasus dormancy buds. A total of 7 197 SSR loci were detected which accounted for 15.62% of total unigenes. Dinuclotide repeat were the most abundant SSR type with a frequency of 58.65%,followed by trinucleotide repeat(34.72%). The availability and polymorphism of selected 8 primers were checked by using 24 cherry accessions. The results showed that average number of effective alleles was 1.73,ranged from a minimum of 1.09(PpSSR8)to a maximum of 2.92(PpSSR2). Shannon’s indexes ranged from 0.202 to 1.290,with an average of 0.755. The observed heterozygosity and expected heterozygosity were from 0.083 to 0.917 and from 0.082 to 0.671,respectively. The average value of observed heterozygosity was 0.391,and of expected heterozygosity was 0.384. The maximum and minimum values of observed heterozygosity,expected heterozygosity and Shannon’s index were observed at locus PpSSR2 and locus PpSSR8,respectively. Genetic relationship of 24 cherry accessions based on the polymorphic primers were not fully in accordance with morphological evidences. Nevertheless,the cherry germplasms were clearly clustered into 5 clades,which indicated an abundant genetic diversity of P. pseudocerasus gathered from Zhejiang Province. Two landraces sampled from Longquan were dramatically different with other cherry accessions. They are genetically more closed to P. serrulata and P. schneideriana. These landraces could be used as parental lines in cherry breeding and were helpful to enlarge the genetic background of cherry cultivars.
Different types of sequences of ORF for GAG in the whole genome of‘Dangshan Suli’(Pyrus pyrifolia White Pear Group)were predicted by bioinformatics methods. A total of 315 GAGsequences were obtained from Ty1-copia retrotransposons. The cluster analysis indicated that there were seven groups,among which,groups one and three had many conserved members. Sequence alignment of all GAG sequences in pear genome showed a high heterogeneity. A total of 34 members were expressed in bagging and bag-removed‘Meirensu’pear fruit,of which,six members(Ppgag16,Ppgag19,Ppgag23,Ppgag27,Ppgag29,and Ppgag30)were highly expressed. In bagging-treated fruit,some members of gag were expressed,suggesting that lightwas not a sole factor to activatethe expression of copia retrotransposons. In bag-removed fruit,the expression of two members(Ppgag5 and Ppgag23)were up-regulated,indicating that some members of the retrotransposons were regulated by light.
The full length cDNA of MrFRK2 was obtained by using RT-PCR and RACE amplification from Chinese bayberry(Myrica rubra Sieb. and Zucc.)‘Biqi’and‘Dongkui’fruit. MrFRK2 was 1 279 bp in full length and encoded a predicted protein of 332 amino acids(ORF length 996 bp),and flanked by 56 nucleotides at the 5′-UTR and 227 nucleotides at the 3′-UTR. The sequence homology comparison showed that,MrFRK2 containedthree conserved regions inherent to the pfkB family of carbohydrate kinases,an ATP-binding domain,and two substrate recognition domains. Phylogenetic analysis indicated that MrFRK2 shared the highest similarity with PtFRK2 with 85% homology. Furthermore,the changes of MrFRK2 expression,fructose and sucrose content during fruit ripening were investigated by q-PCR and HPLC analysis. Meanwhile,Dongkui fruit also experienced higher levels of fructose and lower sucrose-to-fructose ratio than in Biqi fruit. Correlation analysis showed fructose content was significantly negative correlation with the expression of MrFRK2 during ripening. Therefore,our results suggested that the higher fructose content in Dongkui fruit might be associated with lower levels of MrFRK2 expression,thereby increased the sucrose-to-fructose ratio in Biqi fruit during ripening.
To screen key genes involved in the differentiation and development of flowers in chestnut (Castanea mollissima Blume),17 MADS-box genes were identified from the differentially expressed genes(DEGs)between female and male flowers of the mixed inflorescence by RNA-seq analysis. Gene structure analysis showed that two genes were Ⅰ-type MADS box genes as lack of semi-conserved K motif,while the other fifteen genes belonged to Ⅱ-type MADS-box group. And phylogenetic analysis demonstrated that among the 13 MADS-box genes with complete open reading frames(ORFs),there were two B-class genes,one C/D-class gene and three E-class genes based on the ABCDE model hypothesis. Furthermore,quantitative real time PCR was used to investigate the expression profiling of these genes in young leaves,male inflorescences at early stage,male and female flowers in the mixed inflorescence. And for one thing the results confirmed that these genes were DEGs(fold change > 2),for another it indicated that some of these MADS-box genes showed very similar expression pattern. And it assumed that these MADS-box genes might play important roles in the differentiation and development of flowers in chestnut.
A reproducible protocol for somatic embryogenesis was established for the ornamental ginger Hedychium coronarium. Immature filaments were cultured on Murashige and Skoog(MS)medium supplemented with 4 mg · L-1 2,4-D,4 mg · L-1 NAA,1 mg · L-1 6-BA. After culture for 180 days,three types of callus were obtained,type Ⅰcallus were light yellow and friable with tightly packed cell contained dense cytoplasm that appeared to be embryogenic;type Ⅱ callus were white with long,highly vacuolated cells that appeared to be non-embryogenic;type Ⅲ callus were yellow with large,less packed cells contained dense cytoplasm that appeared to between embryogenic and non-embryogenic. Medium containing 1 mg · L-1 2,4-D,0.25 mg · L-1 NAA,0.25 mg · L-1 6-BA was suitable for embryogneic callus proliferation. Embryogenic calli were cultured on somatic embryo induction medium containing MS basal salts,vitamin B5,100 mg · L-1 glutamine,230 mg · L-1 proline,100 mg · L-1 malt extract,0.25 mg · L-1 NAA,0.5 mg · L-1 TDZ,45 g · L-1 sucrose and 7 g · L-1 agar for 20 days,then transferred on MS medium free hormone for 30 days. About 50–60 somatic embryos were induced from per gram callus. Germination percentage of 85% were observed in germination medium,which consisted of MS basal salts,0.2 mg · L-1 NAA and 0.5 mg · L-1 6-BA. Regenerated plantlets with normal shoot and root were developed after 30 d on half strength MS medium with 1 g · L-1 active charcoal. Well rooted plantlets were successfully acclimatized with a survival rate of 90% and grew vigorously. The chromosome number of 100 regenerated plants showed a wide range of variation,89 of them were diploid(2n = 2x = 34),3 were triploid(2n = 3x = 51),6 were tetraploid(2n = 4x = 68)and 2 were hexaploid(2n = 6x = 102).
In order to fast by detect Citrus vein enation virus(CVEV)with sensitivity and specificity,a Reverse Transcription Quantitative Polymerase Chain Reaction(RT-qPCR)assay was developed with selective primer pairs(EVqF4/EVqR4)and optimized conditions. Using the method,only sample infected with CVEV showed positive,while those with other five citrus pathogens were negative. The sensitivity of the method was 100 times higher than the conventional RT-PCR. There was a good linear(R2 = 0.992)relationship between the threshold cycle and CVEV template concentration,while the amplification efficiency of the RT-qPCR was 101.8%. The coefficients of variation of the intra- and inter-assay were both within 2.85%,indicating a good reproducibility of the method. The results also showed that CVEV was uneven distributed in sour orange plants and hybrid citrus plants,and the amount of the virus in roots was the highest(162.52 and 45.32 copies · ng-1 RNA,respectively),followed by that in barks and leaves.
‘Tangxiahong’is a new litchi cultivar selected from offspring plants of mutated individual seedlings in Guangdong. The fruit is short heart-shaped with scarlet skin. The flesh tastes sweet,soft and smooth,which exhibits strong aroma. The yield of third years of top grafting was 14.685 t · hm-2. The average weight of single fruit,edible portion and content of soluble solids were 27.1 g,76.4% and 18.5% respectively.‘Tangxiahong’is a promising cultivar for its rough management,easy to flower as well as high and stable yield.
‘Zibao’is a bud mutation of‘Qingpi’fig(Ficus carica L.)which is one of the most important cultivars growing in China. The mutation demonstrated stable phenotype after continuous asexual reproduction. The main crop syconia of‘Zibao’are 33.6 mm in mean diameter,33.4 g in average weight,of purple skin and deep red pulp. The fig is of very high intrinsic quality,the flesh is luscious and juicy with attractive aroma,the average soluble solids content is 18.5°Brix.
‘Jin Fanqie 10’is a new tomato hybrid which was developed by crossing‘Y33-2’as a female parent with‘T5-1’as a male parent. It is middle ripening and indeterminate growth type with strong growing vigor,dark green leaves,high resistance to ToMV,resistance to CMV,resistance to leaf mold. Its fruit in mature is subglobose,slick,bright pink,and less deformed. It has good quality,good marketability,long storability. Single fruit weight ranges from 200 to 300 g. The marketable yield is more than 110 t · hm-2. It can be well cultivated in greenhouses in Shanxi Province.
Cucumber hybrid cultivar‘Jinyou 409’was released by integrating haploid technology and conventional breeding methods. The plant is of strong growth vigor. The fruit is dark green and glossy,short neck,about 36 cm in length,and of high marketable quality. It is resistant to downy mildew,powdery mildew,Fusarium wilt,and CMV. It can yield 90 t · hm-2 in open field cultivation in spring. It is suitable for open field cultivation.
‘Beautiful Lotus’is a new cultivar selected from the superior individuals of the blue and white Iris sanguinea,and it has long-green period,light purple flower color. Plant height is about 90 cm and stem height is 83.4 cm in blooming period,the crown width is about 40–55 cm after planted two years,and every plant has about 18–30 scape. Flower diameter is about 12 cm. The single scape have about 2–3 flowers. The continuous flowering is about 20 days. It’s suitable to be applied in the north of China.