In order to study the regulation function of MdBZR1 on apple plant height,a BR signal transcription factorgene BRASSINAZOLE-RESISTANT 1(BZR1)was cloned from apple dwarf rootstock Malling 9(M9),which was named as MdBZR1. MdBZR1contains an open reading frame(888 bp)and encodes a protein of 295 amino acids. Bioinformatics analysis showed that MdBZR1 shared five conserved domains:nuclear localization signal(NLS)(MARRKPSWRERENNRRTERRR),nitrogen terminal domain(N)(NLPKHCDNNEVLKALCLQAGWTVEDDGT),BIN2 phosphorylation domain (STKISPY
The apple α-farnesene synthase gene(MdAFS)was cloned by combination of the homologous clone and PCR technology from apple(Malus × domestica‘White Winter Pearmain’). By using chloroplast transit peptide(pla)of the AtCor15A gene from Arabidopsis thaliana,we targeted MdAFS to the chloroplast of Arabidopsis to generate pla + MdAFS transgenic lines. Transgenic Arabidopsis was generated by Agrobacterium-mediated genetic transformation. The subcellular localization of the pla-MdAFS-GFP fusion fluorescent protein revealed that the MdAFS protein was localized in chloroplast. The quantification of photosynthesis-related isoprenoid metabolites in all plants at the rosette and flowering stages showed that chlorophyll a,chlorophyll b,total chlorophyll,and carotenoid were higher in the transgenic lines as compared to wild-type control. The expressions of terpenoid biosynthetic genes were investigated in different development stages by qRT-PCR. Especially,at the flowering stage,the expression of several terpenoid biosynthetic genes significantly increased in the transgenic lines. Gas chromatography mass spectrometry(GC–MS)analysis of volatile extracts from the transgenic plants revealed the biosynthesis of several novel sesquiterpenes,including (E,E)-α-farnesene. These results indicated that overexpressing MdAFS in chloroplast of Arabidopsis could significantly affect the reorientation of terpene metabolic pathway,and increase the transgenic plant emitting terpene volatiles.
In order to explore the mechanism of blue light promoting carotenoids accumulation in postharvest peaches(‘Jinli’,yellow-flesh),PpSGR1 and PpSGRL were identified based on peach genome and analyzed by bioinformatics. Furthermore,the expression profile of PpSGR1 and PpSGRL in different tissues and developmental fruits and the effects of blue light treatment(40 μmol · m-2 · s-1) on PpSGR1 and PpSGRL expression were investigated in‘Jinli’peach fruit during storage. Results showed that PpSGR1 possessed cysteine-rich motif and belonged to STAY GREEN subfamily,while PpSGRL belonged to STAY GREEN-like subfamily due to the mutation of cysteine-rich motif in C-terminal. PpSGR1 and PpSGRL both contained a chloroplast transit peptide in their N-terminal,and shared high similarity homology with Prunus mume. Expression pattern of PpSGR1 and PpSGRL differed significantly in peach
We used eight-year-old‘Zhongyoutao 4’nectarine as the material and then treated dormant peach branches with hormone. The results showed that ABA delayed bud germination,GA promoted germination and precocious flowering,ABA and GA had the adverse effect on the germination of peach buds. We obtained at least 3 homozygous lines with overexpression of ABA 8’-hydroxylase gene which were key enzyme in ABA catabolism in peach such as PpeCYP707A1,PpeCYP707A2,PpeCYP707A3 in Arabidopsis thaliana by transgenic technique. The results of RT-PCR showed that the expression of target gene in overexpression lines reached 23.86 times,7.37 times and 3.23 times,respectively. The Arabidopsis of overexpression lines showed slow growth,delayed bolting even flowering,which PPECYP707A1 slowest growth and flowering the latest. Relative conductivity and active oxygen analysis showed that seed germination and seedling growth of transgenic Arabidopsis were not sensitive to ABA and the damage degree was lower compared to wild type. Low temperature(10 ℃)treatment results showed that the relative growth rate of transgenic Arabidopsis with overexpressing PpeCYP707As was higher than that of wild type. It is speculated that the overexpression of CYP707As gene may positively regulate the growth and development of Arabidopsis under low temperature condition.
Taking the hickory(Carya cathayensis Sarg.)embryo as the test material,five periods(75,85,100,109 and 127 days after pollination)of hickory embryo which are critical for the oil synthesis and integration are measured by gas chromatography to study the law of hickory oil. Additionally,Stearoyl- ACP desaturase genes(CcSAD-1,CcSAD-2,CcSAD-3)and fatty acid desaturase genes(CcFAD2,CcFAD3-1,CcFAD3-2,CcFAD3-3,CcFAD6,CcFAD7,CcFAD8-1,CcFAD8-2,CcFAD8-3)were studied by quantitative real-time PCR. The results revealed that hickory oil mainly composed of palmitic acid(C16︰0),stearic acid(C18︰0),oleic acid(C18︰1),linoleic acid(C18︰2)and linolenic acid(C18︰3);hickory is a high oleic species. In addition,quantitative results showed that with the increase of Stearoyl- ACP desaturase genes expression,the oleic acid content in hickory oil increased and reached dynamic balance. Gene expression profiling and lipid analysis in hickory fruit during development identify CcSAD-3 as the main gene responsible for the oleic acid content in hickory oil. Meanwhile,with the increase of oleic acid content in the hickory oil,the expression of CcFADs increased,and the proportion of unsaturated fatty acids in hickory oil increased.
For the scarcity of persimmon male resources,the systematical collection and arrangement of the male resources as well as the clarity of genetic diversity of these resources and the genetic relationship among them together with other main cultivars and closely related species are essential for the efficient utilization of male resources. In this study,more than 90 wild male persimmon germplasm resources were found in Hubei,Guangxi,Jiangsu provinces,etc.,during the survey of wild persimmon germplasm resources nationwide. Subsequently,15 SSR primers were used to analyse the genetic diversity of 47 male persimmon resources,2 andromonoecy resources and 11 Chinese monoecious resources. Sixty-seven female resources,38 Japanese resources(including 25 monoecious resources)and 28 closely related species were used as reference out-group. The results indicated that a total of 144 alleles and 316 genotypes were obtained from 15 loci,with a mean value of 9.60 alleles and 21.07 genotypes per locus. ssrDK11/DQ097479 was the most informative locus. The geographical distribution of male persimmon resources indicated that they were common in wild comfortable remote area. The male resources originated in Mulan Mountain,Hubei Province and in Shaanxi Province showed the highest and the lowest genetic diversity,respectively. The 193 resources were divided into 16 and 12 clusters by UPGMA and ME cluster analysis,respectively. Similar general laws were found according to the two cluster analysis results:(1)a genetic isolation existed among wild male resources from different areas;(2)the wild male resources in Mulan Mountain,Hubei Province could be genetically divided into two clusters,one had a near relationship with Jiangxi,Jiangsu,Hunan wild persimmons as well as main astringent cultivars such as‘Huojing’and monoecious astringent cultivars such as‘Yaoxian Wuhuashi’,while the other was closer to Yunjia Mountain male resources and Luotian non-astringent persimmons(including‘Tianbaogai’);(3)most Japanese resources could be classified into a same cluster exclusively.
In order to investigate the effect of tomato rootstock,scion and their potential interaction on chilling tolerance,we grafted chilling-sensitive tomato cultivar‘060911’(S)and chilling-resistant tomato cultivar‘060112’(R). We then selectively removed rootstock or scion from the grafted seedlings RS/RS,and incubated the newly formed 9 kinds of seedlings,i.e.,RS/RS,RS/R,RS/S,R/RS,S/RS,S/R,R/S,R/R,S/S in the growth chamber with light intensity of 22 klx and photoperiod 12 h/12 h. The seedlings were treated with low temperature at 10 ℃/3 ℃ for 9 days and recovered at 25 ℃/15 ℃ for 3 days. Chilling injury index,root activity,electrolyte leakage and osmotic adjustment substances were measured after the chilling treatment and after the recovery. Grafted seedlings behaved similarly in root activity,electrolyte leakage of root and leaf,soluble sugar and proline content before the chilling treatment,but showed significant difference after the chilling treatment and after the recovery. Among these 9 kinds of grafted seedlings,S/S seedlings displayed the highest while R/R seedlings displayed the lowest chilling injury index. Meanwhile,R/R seedlings showed the lowest electrolyte leakage,and the highest content of soluble sugar and proline. Meanwhile,R/R seedlings showed the lowest electrolyte leakage,and the highest content of soluble sugar and proline. While R/RS,RS/R,RS/S,S/RS seedlings were intermediate between R/R and S/S regarding the chilling injury index,electrolyte leakage,soluble sugar and proline content. Results indicated that both rootstock and scion are important for chilling tolerance. We further found that rootstock conferred higher chilling resistance than scion,due to the fact that the chilling tolerance of RS/RS was higher than that of S/RS,but lower than R/RS. Moreover,the chilling tolerance of RS-R was significantly higher than that of RS-S,and the P value of interaction was less than 0.01. The results showed that rootstock and scion had significant interaction effect on the chilling tolerance of grafted seedlings.
A 3-year experiment was conducted to investigate the change pattern of daily yield and its response to sunlight duration and air temperature with different phosphorous rates during cucumber growing season under greenhouse condition. The formation of yield peak,the growth period for nutrient use with the highest efficiency and the appropriate phosphate rate were discussed in order to solve the problem of excessive application of phosphorus in greenhouse production in the North China Plain. Three treatments were designed in this experiment,including a non-phosphorus control(P0),a recommended phosphorus rate(P1)and a farmers’ conventional phosphorus rate(P2). Based on the basic soil Olsen-P content,the abundance index of soil phosphorus and the target yield,the phosphorous rate was recommended as P2O5 300 kg · hm-2 in the P1 treatment. Based on the average amount of phosphate applied by farmers planting greenhouse cucumber in Hebei Province,the phosphorus rate of the P2 treatment was designed as P2O5 675 kg · hm-2. The results showed that:(1)quadratic curve was used for describing the daily yield formation during 3 years:the daily yield peak was observed in 97–104 days after transplanting with the accumulated air temperature(8 am,≥ 10 ℃)1 389.4–1 849.6 ℃ and the accumulated sunlight duration 629.0–866.8 h:the average daily temperature was 23–27 ℃ and the estimated active accumulated air temperature was 1 650–2 050 ℃ at this point.(2)Although 56% of phosphate was reduced,the soil phosphorus supply in the P1 treatment was sufficient for yield formation and no significant change was observed in the yield peak and the total yield compared to the P2 treatment. (3)The yield peak was postponed for 16 days and close to the vine removed stage in the P0 treatments compared to the P2 treatment in 2009,which indicated that continuous non-phosphorus application leaded to the delay of the fruit maturation when high yield was formed. In conclusion,for winter-spring cucumber,the growing stage for nutrient utilization with the highest efficiency was considered from the end of May to the early June,because of forming yield peak during this period. Sufficient fertilizer and water application would be helpful to improve nutrient use efficiency during this period. For the greenhouse with the similar test condition,applying P2O5 300 kg · hm-2 for the target yield of 170 t · hm-2,would guarantee no change of the yield peak and the total yield and to reduce over 50% phosphate compared to the farmers’ conventional management.
Through the field observation and experimental study,the outcrossing index(OCI),pollen vitality and stigma receptivity,the ratio of pollen and ovule(P/O),artificial pollination and bagging were investigated to understand the flower structure and reproduction system in Freesia hybrida. The results showed that the stamen of Freesia hybrida was closed to the lateral side of the style and about 2 mm shorter than the stigma. The relative position between the stamen and gynoecium was unchanged during the whole flowering stage. In addition,the stamen and gynoecium of Freesia hybrida mature at different times,the highest period of receptivity stigma was not synchronized with the peak stage of pollen viability. The OCI and P/O are 5 and 1 614,respectively. Taking the results of the artificial pollination experiment together,it indicated that reproduction system of Freesia hybrida was facultative outcrossing with outcrossing as the main breeding form and self-pollination as auxiliary form. Artificial pollination on fruit set rate was higher than the natural pollination.
To provide theoretical basis for genetic evolution and diversity of Osmanthus fragrans,reverse transcriptase(RT)sequences of Ty1-copia and Ty3-gypsy-like retrotransposon were cloned and analyzed in the genome. Using degenerate oligonucleotide primers corresponding to conserved domains of Ty1-copia and Ty3-gypsy LTR retrotransposon RT sequence,fragments of 260 bp and 430 bp were amplified by polymerase chain reaction(PCR)from genomic DNA of leaves,respectively. Then,the products were purified and sub-cloned to pMD-19-T vector. Subsequently,49 RT sequences of Ty1-copia-like retrotransposon and 20 of Ty3-gypsy-like retrotransposon were obtained through sequencing and related bioinformatics analysis. RT sequences of Ty1-copia-like retrotransposon were classified into four groups after cluster and alignment analyses of their nucleotide sequences. The length of the nucleotide sequences varied from 255 to 271 bp,and homology ranged from 32.0% to 96.9%. When RT nucleotide sequences translated into amino acids,two sequences showed frameshift mutation,and sixteen sequences presented stop codon mutation. Four groups were clustered by alignment analyses of RT nucleotide sequences of Ty3-gypsy-like retrotransposon. The length of the nucleotide sequences varied from 408 to 449 bp,and homology ranged from 54.7% to 98.8%. The deduction of amino acids revealed that 8 sequences presented frameshift mutation and 10 sequences presented stop codon mutation. Phylogenic trees were constructed based on the amino acid sequences from other species,indicating that Osmanthus fragrans had homologies with Prunus mume,Prunus salicina as well as Ziziphus jujuba. The RT nucleotide sequence of Ty1-copia and Ty3-gypsy retrotransposon were highly heterogeneous. Phylogenetic analysis demonstrated that Osmanthus fragrans showed a common origin with other species,and a horizontal transmission of retrotransposons occurred among different species in the process of evolution.
In this study,a nucleotide sequence in length of 1 999 bp was cloned from tea plant(Camellia sinensis),termed as CsSULTR3.1(GenBank accession No. KY963793). Bioinformatics analysis showed that the sequence contained 1 980 bp Open Reading Frame(ORF) and encoded 659 amino acid residues. BlastX and phylogenetic analysis indicated that CsSULTR3.1 shared the highest identity(86%)with SULTR3.1 in Vitis vinifera. In addition,CsSULTR3.1 was speculated as a non-secretory protein with 10 transmembrane domains,72.39 kD molecular weight and 7.61 theoretical isoelectric point. Subcellular localization using GFP assays validated that CsSULTR3.1 was located in cytoplasm. Results of real-time PCR analysis showed that CsSULTR3.1 could be detected in all tested tissues of tea plant and was highly expressed in stems. Furthermore,CsSULTR3.1 could be induced by the treatments of Na2SO4 and Na2SeO4,respectively. Under 60 mg · L-1 sulfate condition,the expression of CsSULTR3.1 gradually raised after 12 h,and reached maximum at 48 h. However,a significant expression decrease was detected within 12 h under 240 mg · L-1 sulfate treatment,and an expression stimulation was observed after 24 h treatment. In comparison,the expression of CsSULTR3.1 showed same change tendency either under high or low selenate treatment condition,briefly decreased firstly,then increased with a peak at 48 h and followed by another decrease.
Rhizosphere microorganisms have great effects on plant growth and development. The exudates of plants can also affect the distribution and species of microorganisms in rhizosphere. In this study,a high throughput sequencing technology was used to clarify the effects of MdSOS2L1 transgenic Gala apple plants on rhizosphere microorganisms. The results showed that there were great differences in the abundance and diversity of rhizosphere microorganisms among the MdSOS2L1 transgenic apple plants in compared with wild type apple plants. Among them,the rhizosphere of prokaryote community varied obviously in phylum level. The abundance of Acidobacteria in transgenic apple rhizosphere was higher than that in wild type apple plants. The abundance of Cyanobacteria and Actinobacteria was lower in transgenic apple rhizosphere microorganisms. Meanwhile,the pH value of rhizosphere soil of transgenic plants was lower than that of wild type plants. And the contents of oxalic acid,citric acid and L-malate in transgenic plants root were higher than that in wild type plants rhizosphere soil,but no significant differences occurred among the bulk soil between these two types of apple plants. Above of all,it is suggested that the MdSOS2L1-mediatedsecretion of organic acids contributes to the difference in rhizosphere microorganisms.
Celery(Apium graveolens L.)is an important vegetable in Apiaceae family,with excellent source of carotenoid. Lycopene epsilon cyclase is a key enzyme in the pathway of carotenoid biosynthesis. Here,the AgLCYE gene encoding lycopene epsilon cyclase was cloned from A. graveolens‘Jinnan Shiqin’. The sequences of AgLCYE gene contained an open reading frame with the length of 1 590 bp,which encoded 529 amino acids. Multiple alignments of the AgLCYE amino acid sequences from celery and other different plants showed a homology of 77.68%. The homology of LCYE was up to 98.07% between celery and carrot. Phylogenetic tree analysis showed that the LCYE proteins were conserved. The physicochemical property analysis showed that AgLCYE belonged to hydrophilic protein. There were 9 alpha helices and 18 beta folds in the predicted tertiary structure of the protein. The results of disordered analysis showed that the amino acid sequence of AgLCYE protein had four disorder regions. The contents of lutein and α-carotene of celery leaves in different growth stages(30,45,and 60 d)were determined by UPLC. The peak of lutein content was detected at 30 d. The lowest content of lutein was observed in 45 d. There was no significant difference between 45 d and 60 d. The lutein content in leaf blade was higher than that in petiole of celery. The presences of α-carotene were not detected in leaf blade and petiole of celery,respectively. The expression levels of AgLCYE gene in celery were detected by quantitative real-time PCR. In leaf blade,the relative expression levels of AgLCYE gene increased gradually over the process of celery growth and development. On the contrary,the relative expression levels of AgLCYE gene decreased in petiole.
The effects of foliar application of silicon on growth,yield,accumulation of sugar and vitamin C in fruit and activities and gene expression of the related enzymes of cucumber were studied. The results showed that foliar application of silicon significantly promoted the growth and yield of cucumber plants,and increased activities of neutral invertase(NI),sucrose phosphate synthase(SPS),sucrose synthase(SS)and galactose dehydrogenase(GaILDH)and contents of fructose,glucose and vitamin C in fruits of cucumber. Among all treatments,foliar application of 1.420 g · L-1 monosilicic acid showed the most significant effect. Compared to the control,contents of fructose,glucose and vitamin C of the fruits increased by 13.79%,18.08% and 75.11%,respectively. Real-time quantitative PCR results indicated that the genes expression of neutral invertase-3(NI-3),sucrose phosphate synthase(SPS)and galactose dehydrogenase(GaILDH)were significantly increased in fruits under 1.420 g · L-1 monosilicic acid treatment.
In Arabidopsis,WIN1/SHINE1 gene has the function of regulating wax synthesis and metabolism. In this study,a homologous gene of WIN1/SHINE1 was cloned from cucumber and named CsWIN1. The results of qRT-PCR showed that CsWIN1 expressed at high levels in the leaves and young fruits of cucumber. Subcellular localization analysis suggested that the CsWIN1 protein is located in the nucleus. The CsWIN1 overexpression in the transgenic plants of Arabidopsis thaliana caused a similar phenotype to WIN1/SHINE1 overexpressing lines of Arabidopsis. The expression analysis of CsWIN1 transgenic lines indicated that the expressions of the genes related to wax synthesis have been changed significantly. These results suggested that the function of CsWIN1 is conserved in the regulation of epidermal wax synthesis through regulating the expression of downstream genes related to wax synthesis to affect wax biosynthesis and metabolism.
In this study,results of liquid chromatography analysis showed sucrose content in leaves of cucumber seedlings under shading treatment was slightly higher than control. While galactinol,raffinose and stachyose contents of shading leaves were lower than that of control. The change of galactinol synthase activity was consistent with galactinol synthase genes expression level,which of shading leaves were lower than control;the activity of raffinose synthase is not significantly different between shading leaves and control,while the transcription level of raffinose synthase genes under shading condition was significantly lower than control;the activity of stachyose synthase of shading leaves was lower than control,and the expression level of stachyose synthase genes was consistent with the activity. In addition,the expression level of SNF1-related protein kinase 1 gene(SnRK1)of shading leaves was higher than control at 12:00 am of first day under 6 h shading treatment,which indicated that SnRK1 occurred in response of shading stress. Together,these results suggested that under shading stress the gene expression and activities of galactinol synthase,stachyose synthase,and SnRK1 in cucumber were regulated to maintain the content of sucrose,provide the energy for growth,and ensure the survival of leaves.
Crabapple combines charming flowers,colorful leaves and attractive fruits in one body. It is a kind of important ornamental plant in northern temperate regions. A systematic review on the progress of classification of crabapple cultivars was conducted. It included the traceability and organ evolution of cultivars,brief history and current situation of classification. Simultaneously,problems existed and corresponding advice were pointed out. The aim of this review was to provide comprehensive and valuable information for further research on the evaluation of germplasm,breeding,genetic diversity,genetic relationship and industrialization.
‘Shuangdongmi’,a new very late-ripening peach cultivar,was derived from the cross of ‘Dongxuemi’בQingzhoumi’. The average fruit weight is 148 g. The fruit is round with a dark purple red pericarp. The fruit flesh is green and white,freestone,succulent crisp with sweet flavor. It has small stone and no dehiscent fruit. The soluble solids content in fruit juice is 14.8%,and the flesh firmness is 11.6 kg · cm-2. It has good character for transportation and storage. The fruit ripens in early November,and the fruit development period is about 210 days. This cultivar has good ability of early fruiting and high productivity. Four-year-old tree yields up to 52 500 kg · hm-2.
‘Nongda Yuxiang’is a new kiwifruit cultivar selected from offsprings of‘Xuguan’. The fruit is long cylinder-shaped,light brown peel,with scabrous tomentum,yellow-green pulp,little core,juicy flesh. The average fruit weight is 110 g. Total sugar content is 11.2%,total acid conent is 1.04%,vitamin C content is 2.52 mg · g-1 and soluble solids content of soft-ripe stage is 18.8%. It has good quality and relatively long storage life. The ripening date is in the early October.
‘Yayou 7’is a new Carya cathayensis cultivar selected from wild resources in Jiande,Zhejiang. The fruit is larger,the average weight is 4.96 g. The average height of fruit and width are 23.90 and 20.43 mm. The fruit shape index is 1.17. The kernel percent is 46.60%,fat content of nutlets is 64.00%,the protein content is 59.80 mg · g-1,the potassium content is 2 733.7 μg · g-1,the calcium content is 1 621.5 μg · g-1,the multiple characteristic is excellent.‘Yayou 7’has the properties of early bearing and high-yield. The average yield of seven-year-old tree of‘Yayou 7’is 1 458.3 kg · hm-2.
‘Ximi 5’is a new hybrid of muskmelon cultivar. Fruit is a high round. The average fruit weight is 1 200–1 500 g. The pericarp is light grey-green with dense and thick netty streaks. The fruit fresh is light green and crisp,4 cm thick. The average soluble solids content is 11.8%. It is medium ripening muskmelon. The whole fruit growth period is 100–105 d and fruit developing period is 42–45 d. The yield is 40 000–48 000 kg · hm-2. ‘Ximi 5’is comparably tolerant low temperature and low light. It is resistant to downy mildew,powdery mildew and tolerant to storage.
‘Dicai’,a new Phalaenopsis cultivar with purple flower,is derived from the cross of‘Jiuhbao Red Rose’and‘Red Dragon’. The flower diameter is 9.2 cm,and the whole inflorescence has 7.6 flowers which have good arrangement. This cultivar shows good characteristics as orderly flowering and bright flowers,etc.