A gene named MdSUT2(MDP0000277235)was cloned from‘Gala’apple(Malus × domestica Borkh.). Sequence analysis showed that this gene contained 1 704 bp nucleotides,and encoded a protein of 566 amino acids. Overexpression vector of MdSUT2 was constructed and transformed into Arabidopsis seedlings via Agrobacterium-mediated method. Semi RT-PCR analysis indicated that MdSUT2 was genetically transformed into Arabidopsis. In addition,the transient expression vectors were constructed and transformed into apple fruits. We found that overexpression of MdSUT2 in apple fruits significantly increased the anthocyanin content,whereas suppression of MdSUT2 decreased the anthocyanin content.
‘Nagafu 2’apple trees were sprayed with 1.5% and 3% glucose solutions at 25 days and 30 days after full bloom,respectively. Bud growth,endogenous sugar levels,starch content and activities of related enzymes in terminal spur buds were examined. Genes associated with sugar metabolism and flowering were studied by quantitative real-time PCR. The results revealed that glucose treatments increased bud size and fresh weight. The contents of sucrose,glucose and sorbitol were significantly higher than in the control during early stage of floweral bud differentiation,while starch increased generally,but the content of fructose significantly decreased during the whole period. The activities of sorbitol dehydrogenase,sorbitol oxidase,synthesis direction of sucrose synthase and sucrose phosphate synthase increased at different stages,while acid and neutral invertases,decomposition direction of sucrose synthase decreased. Carbohydrate accumulation in leaves was evident,and the activities of enzymes involved in sugar metabolism also changed similarly. Genes associated with sugar metabolism and two MdTPS genes in terminal buds all responded to glucose treatment. Furthermore,flowering genes like MdFT 1,MdFD,MdLFY,MdSOC 1,and MdAP 1 were significantly up-regulated,while the suppressor MdTFL 1 was down-regulated from beginning of floral induction.
The 2 211 bp CDS sequence of PbTMT4 gene(Pbr032130.1)and the promoter sequence of 1 220 bp upstream of the transcription initiation site were obtained using Pyrus bretschneideri‘Dangshan Suli’as the material based on the pear genome database. When the PbTMT4 gene was introduced into Arabidopsis by Agrobacterium-mediated method,the growth rate of transgenic Arabidopsis was faster,and the flowering time was earlier than the wild type. At the same time,the amount of sugar accumulation in transgenic Arabidopsis was significantly higher than that in wild type. Bioinformatics analysis indicated that the promoter contains multiple cis-acting elements associated with stress response,hormone signals and light signals. To further analyze the promoter function of PbTMT4 gene,a promoter-reporter vector was constructed and transformed into Arabidopsis. Analysis of T3 transgenic Arabidopsis tissues with histochemical staining and semi-quantitative analysis showed that GUS gene was expressed in roots,stems,leaves,flowers and fruit pod. And sodium chloride(NaCl),drought,gibberellin(GA3),methyl jasmonate(MeJA)and light intensity can induce the transcription level of GUS gene in Arabidopsis to some extent. Stress treatment revealed that the PbTMT4 gene overexpression transgenic line had less damage than the wild type plants. The results of the study preliminarily suggest that PbTMT4 gene can promote the development of transgenic plants and increase sugar accumulation,and may play an important regulatory role in the abiotic stress.
The levels of primary metabolite in the fruit pulp of somatic hybrid [Citrus sinensis Osbeck‘Newhall’+ Citrus limon(L.)Burm. F.‘Eureka’] and both parental species were quantitatively evaluated by GC–MS(gas chromatography-mass spectrometry)and flavonoid contents were investigated by LC–MS(Liquid chromatography-mass spectrometry). The results indicated the allotetraploid was closer to the lemon parent in the accumulation of primary metabolites. The levels of malic acid,citric acid and four fatty acids were higher than those of the diploid parents,while the contents of most amino acids in the allotetraploid showed an additive manner. Additionally,a total of 48 flavonoids were detected in the allotetraploid. The contents of 33 flavonoids were intermediate between both parents. Meanwhile,the levels of 15 flavonoids were transgressive. The allotetraploid was closer to the lemon parent in flavonoid composition.
The ζ-carotene desaturase(ZDS)gene was selected as the target gene to establish the CRISPR/Cas9 genome editing system in Chinese kale. The target site,near the 5′ end of the coding region of the BoaZDS,was selected,and the CRISPR/Cas9 expression vector was constructed. As a result,a total of 19 transgenic plants were obtained through Agrobacterium-mediated genetic transformation. Sanger sequencing analysis revealed that 13 of them were mutated. The mutation efficiency was 68.42%,and all mutants showed a clear albino phenotype.
To understand the mechanism of bud mutation in Malus × domestica Borkh.‘Fuji’,DNA methylation patterns and epigenetic variations were identified using MSAP(Methylation Sensitive Amplified Polymorphism)molecular marker analysis and UPGMA cluster analysis in a population of 35 ‘Fuji’bud sports lines(Red Fuji). Results indicated that:(1) MSAP amplication pattern varied among different bud sports lines. Total methylation was ranged from 27.90% to 36.16%,and 32.87% in average. Inner-methylation of double-stranded DNA was the main pattern in a primary‘Fuji’cultivar and its bud sports lines. (2) The CCGG sites remained primary methylation patterns during the occurrence of bud sports together with‘Fuji’. (3) In total,12 methylation variation patterns were detected from 68.57% Red Fuji lines. The frequence of hypomethylation was significantly higher than that of hypermethylation. Moreover,CG hypomethylation was significantly higher than that of CHG(P < 0.01). (4) Average similarity coefficience among 36 bud sports lines was 0.89,ranging from 0.79–0.92. In the UPGMA dendrogram,the primary Fuji cultivar fell out of the group of bud sports lines. Most recently bud spot lines had tendency to cluster together;typeⅠ(flushed-skin color pattern)and typeⅡ(striped-skin color pattern)bud sports lines were mixed into differents. In summary,DNA methylation and demethylation happened at the same time,demethylation was the main methylation pattern. CG hypomethylation plays an important role in the occurrence of Red Fuji. Our results would be great help in future epigenetic study of‘Fuji’sports lines,and CG hypomethylation would be as an emphysis to explore the mechanism of sports line occurence.
The distribution of simple sequence repeat(SSR)was studied in 1 431 scaffolds of Chinese bayberry(Myrica rubar Sieb. et Zucc.)genome. There were 43 842 SSR loci identified totally. And 194 kinds of repeating motifs that distributed in 623 scaffolds were discovered. The number and frequency of dinucleotide SSR loci were 36 383 and 82.99%. The frequency of AG/CT in dinucleotide was 55.70%,and in all SSR loci was 46.23%. It was indicated that AG/CT was the most common repeat motifs in Chinese bayberry genome. The number of trinucleotide SSR loci was 6 115,accounting for 13.95%. The number of tetranucleotide,pentanucleotide and hexanucleotide were 827,245 and 272 with the frequency 1.89%,0.56% and 0.62%,respectively. Meanwhile,59 pairs of Genomic SSR markers(gSSRs)were developed. After polymorphism analysis,26 gSSRs were highly polymorphic markers and 33 gSSRs were moderately polymorphic markers. In this study,the clustering analysis among the advanced selection Zaoxian 856 and the other 13 main cultivars were identified by all 59 gSSRs. According to the result,14 Chinese bayberry materials were clustered into two groups at the similarity coefficient of 0.56. It was illustrated that Zaoxian 856 was different from the other 13 varieties at the DNA level,and the similarity coefficient between Zaoxian 856 and control‘Zaose’was 0.86.
‘Jinyudian’is a new cultivar which is bred from the natural variant plants of Luculia pinceana. The cultivar is perennial evergreen shrub with erect plant type. Its thin leathery leaf is ovate in shape with leaf index 1.81. The inflorescence type is cyme and the flower type is long-style. The viridescent sepal is half-reversal in shape at anthesis and its abaxial surface is glabrous. The corolla tube is (3.52 ± 0.13)cm in length. The corolla diameter is(2.87 ± 0.27)cm with light pink in color. There are tiny flake attachments between corolla lobes. The cultivar has great ornamental value,which can be planted for domestic decoration and landscape designment widely.