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To understand the relationship between fruit cracking and gene expression patterns, we identified two expansin genes from litchi (Litchi chinensis Sonn.) fruit and then examined their expression profiles in pericarp and aril at different stages of fruit development, using the cracking-resistant cultivar Huaizhi and the cracking-susceptible cultivar Nuomici. Two full-length cDNAs of 1087 and 1010 base pairs encoding expansin, named LcExp1 and LcExp2, were isolated from expanding fruit using RT-PCR and RACE-PCR (rapid amplification of cDNA ends) methods. LcExp1 mRNA could be detected from the early stage of fruit rapid growth (59 days after anthesis). The LcExp1 mRNA increased and reached to the highest level at the end of growth phase (80 days after anthesis) in pericarp of `Huaizhi', while the mRNA could be detected at the stage of rapid fruit growth, then increased slightly and finally kept remained almost constant in the pericarp of `Nuomici'. Similar accumulation of LcExp2 mRNA was observed in fruit aril of `Nuomici' and `Huaizhi', whereas LcExp2 accumulated only in pericarp of `Huaizhi' but did not appear in pericarp of `Nuomici'. The results indicate that expression of two expansin genes in litchi pericarp are closely associated with fruit growth and cracking.
Cold stress is an important factor that limits grape (Vitis sp.) production around the world. The high expression of osmotically responsive genes 1 (HOS1) protein acts as a repressor of cold-responsive genes in plants. To increase understanding of mechanism regulating cold tolerance in grape, we isolated and characterized a novel HOS1 gene, designated VvHOS1 from ‘Muscat Hamburg’ grapevine (Vitis vinifera). Real-time polymerase chain reaction (PCR) analysis revealed that the expression of VvHOS1 could be induced by the application of exogenous abscisic acid and various abiotic environmental conditions such as low temperature, drought, and salinity. Moreover, VvHOS1 expression could also be induced by cold plus drought conditions (4 °C, 10% polyethylene glycol 6000). In addition, overexpression of VvHOS1 in arabidopsis (Arabidopsis thaliana) decreased the plants’ tolerance to cold, drought, and salt as well as negatively regulated the expression level of two stress-responsive genes, AtRD29A and AtCOR47. The results obtained in this study should help us to elucidate the function of VvHOS1 and understand the cold-responsive pathway in grapevine.
Epimedium species are traditional Chinese medicinal plants as well as potential groundcover and ornamental plants. In this study, genome size and genome structures of Epimedium species were investigated using flow cytometric and fluorescence in situ hybridization (FISH). The nuclear DNA content of Epimedium species ranged from 8.42 pg/2C (8230.7 Mbp) to 9.97 pg/2C (9752.8 Mbp). The pairwise nucleotide diversity (π) of the fragments of the genes for reverse transcriptase (rt) of Ty1-copia retrotransposon within a species of rt fragments ranged from 0.251 to 0.428 in 10 Epimedium species. Phylogenetic analysis of the sequences revealed four major clades with the largest subclade containing 72 sequences of relatively low nucleotide diversity. FISH indicated that Ty1-copia retrotransposons are distributed unevenly along the pachytene chromosomes of E. wushanense and E. sagittatum, mostly associated with the pericentromeric and terminal heterochromatin. The relatively low sequence heterogeneity of Ty1-copia rt sequences implies that the Epimedium genomes have experienced a few relatively large-scale proliferation events of copia elements, which could be one of the major forces resulting in the large genome size of Epimedium species.
A pot experiment with summer cover crops and soil amendments was conducted in two consecutive years to elucidate the effects of these cover crops and soil amendments on `Clemson Spineless 80' okra (Abelmoschus esculentus) yields and biomass production, and the uptake and distribution of soil nutrients and trace elements. The cover crops were sunn hemp (Crotalaria juncea), cowpea (Vigna unguiculata), velvetbean (Mucuna deeringiana), and sorghum sudangrass (Sorghum bicolor × S. bicolor var. sudanense) with fallow as the control. The organic soil amendments were biosolids (sediment from wastewater plants), N-Viro Soil (a mixture of biosolids and coal ash, coal ash (a combustion by-product from power plants), co-compost (a mixture of 3 biosolids: 7 yard waste), and yard waste compost (mainly from leaves and branches of trees and shrubs, and grass clippings) with a soil-incorporated cover crop as the control. As a subsequent vegetable crop, okra was grown after the cover crops, alone or together with the organic soil amendments, had been incorporated. All of the cover crops, except sorghum sudangrass in 2002-03, significantly improved okra fruit yields and the total biomass production (i.e., fruit yields were enhanced by 53% to 62% in 2002-03 and by 28% to 70% in 2003-04). Soil amendments enhanced okra fruit yields from 38.3 to 81.0 g/pot vs. 27.4 g/pot in the control in 2002-03, and from 59.9 to 124.3 g/pot vs. 52.3 g/pot in the control in 2003-04. Both cover crops and soil amendments can substantially improve nutrient uptake and distribution. Among cover crop treatments, sunn hemp showed promising improvement in concentrations of calcium (Ca), zinc (Zn), copper (Cu), iron (Fe), boron (B), and molybdenum (Mo) in fruit; magnesium (Mg), Zn, Cu, and Mo in shoots; and Mo in roots of okra. Among soil amendments, biosolids had a significant influence on most nutrients by increasing the concentrations of Zn, Cu, Fe, and Mo in the fruit; Mg, Zn, Cu, and Mo in the shoot; and Mg, Zn, and Mo in the root. Concentrations of the trace metal cadmium (Cd) were not increased significantly in either okra fruit, shoot, or root by application of these cover crops or soil amendments, but the lead (Pb) concentration was increased in the fruit by application of a high rate (205 g/pot) of biosolids. These results suggest that cover crops and appropriate amounts of soil amendments can be used to improve soil fertility and okra yield without adverse environmental effects or risk of contamination of the fruit. Further field studies will be required to confirm these findings.
Carpetgrass [Axonopus compressus (Sw.) Beauv.] is an important warm-season perennial turfgrass that is widely used in tropical and subtropical areas. The genetic diversity of 63 carpetgrass accessions in China was studied using simple sequence repeat (SSR) markers. Fourteen SSR primer combinations generated a total of 49 distinct bands, 48 (97.96%) of which were polymorphic. The number of observed alleles ranged from 2 to 6, with an average of 3.5. Coefficients of genetic similarity among the accessions ranged from 0.24 to 0.98. Unweighted pair-group method with arithmetic means (UPGMA) clustered the 63 accessions into three groups, and not all samples from the same region belonged to the same group. SSR markers will promote marker-assisted breeding and the assessment of genetic diversity in wild germplasm resources of carpetgrass.
Expansins are extracellular proteins that are involved in cell wall modifications such as cell wall disassembly, cell separation, and cell expansion. Little is known about expansin gene expression during flower development of wintersweet (Chimonanthus praecox). In the present study, an expansin gene, CpEXP1, was isolated from the wintersweet flower cDNA library through random sequencing; this gene encodes a putative protein of 257 amino acids with the essential features conserved, like in other alpha expansins. The CpEXP1 gene exhibited different transcription levels in different tissues and had a significantly higher expression in flowers than other tissues. It is strongly correlated with the development of the flower. The expression of CpEXP1 increased in the flower buds or whole flowers from Stage 1 to 4 and decreased from Stage 5 to 6 during natural opening. Ethephon (an ethylene releaser) treatment promoted cut flower senescence, whereas 1-methylcyclopropene (1-MCP) (an ethylene perception inhibitor) delayed the process of flower wilting. This result is associated with the concomitant lower transcript levels of CpEXP1 in the ethephon-treated samples as well as the steady expression in the 1-MCP-treated samples compared with that in control flowers. The studies show the interesting observation that the expression of an expansin gene CpEXP1 is correlated with the development of Chimonanthus praecox flowers, the upregulation during flower opening vs. the downregulation during senescence.
‘Jincuilei’ is a mutant selected from Lonicera macranthoides Hand.-Mazz. It produces abundant flowers that never open with a chlorogenic acid (CGA) content up to 6.0%. Propagation through rooting or grafting has only a 30% survival rate. This study was undertaken to establish an efficient protocol for rapidly regenerating this mutant. Leaf explants were inoculated on Gamborg's B5 medium supplemented with different concentrations of 6-benzyladenine (BA) and 2,4-dichlorophenozyacetic acid (2,4-D). The optimal combination for callus induction was 4.4 μm BA with 2.26 μm 2,4-D, which resulted in 86.7% of leaf explants producing calluses in 4 weeks. Calluses produced from this optimal medium were cultured on B5 medium containing different concentrations of kinetin (KT) and α-naphthalene acetic acid (NAA). The best formulation for shoot induction was B5 medium containing 0.9 μm KT and 5.4 μm NAA in which 73.4% of cultured calluses produced shoots in 8 weeks, and shoot numbers ranged from three to six per callus piece (1 cm3). Adventitious shoots were cut and rooted in half-strength Murashige and Skoog medium supplemented with 14.8 μm 3-indolebutyric acid. Roots initiated 10 d after culture, and rooting percentages ranged from 98% to 100%. Plantlets grown in a container substrate in a shaded greenhouse had over a 95% survival rate. During the last 6 years, over four million plantlets were regenerated using this established procedure, and there was no somaclonal variation. Fresh and dry weights of 1000 flowers, CGA contents, and dry flower yields of the regenerated plants were not significantly different from those of the stock ‘Jincuilei’ propagated by cutting, indicating that plants regenerated from this established procedure were stable. This established in vitro culture method has led to rapid commercial production of this medicinal plant on more than 1500 ha of production field.
Leaf mold, caused by the fungus Cladosporium fulvum, is a serious disease of tomato. In the current study, the main physiological races of C. fulvum collected from three northeastern provinces of China were identified using a set of identification hosts. The results showed that the prevalent pathogenic physiological races were 1.2.3, 1.3, 3, 1.2.3.4, and 1.2.4. F1, F2, and BC1 tomato plants were obtained by crossing C. fulvum-resistant cultivar 03748 carrying the Cf-6 gene and susceptible cultivar 03036. Three 10-mer oligonucleotide random amplified polymorphic DNA (RAPD) primers and two simple sequence repeat (SSR) primers were selected for the further molecular marking analysis after 210 RAPD primers and 50 SSR primers were screened using the bulked segregate analysis method. The polymorphic DNA bands were amplified among parents, 10 F1 plants, 184 F2 plants including 145 resistant plants and 39 sensitive plants using three RAPD primers and two SSR primers so that three RAPD molecular markers and two SSR molecular markers linked to the Cf-6 loci were identified. Three RAPD markers were linked to the Cf-6 resistant locus separated with 8.7 cM, 20.3 cM, and 33.4 cM. Also, one RAPD codominant marker S374619/559 was found. The locations of the two SSR markers were 12.6 cM and 9.7 cM away from the Cf-6 locus. After cloning and sequencing two specific DNA fragments closely connected to the Cf-6 resistant and susceptible alleles respectively, in the RAPD codominant marker S374619/559 and one codominant sequence characterized amplified region marker S674619/559 was converted from RAPD marker S374619/559. In the RAPD marker S374619/559, the length difference of two specific fragments, 619-bp fragment and 559-bp fragment, is the result of one insertion (60 bp) in the 619-bp fragment. These markers will facilitate the selection of resistant tomato germplasm containing the Cf-6 gene and cloning of Cf-6 to breed new C. fulvum resistant tomato cultivars.
In an effort to mitigate the environmental impact of chemical fertilizers, plant growth-promoting rhizobacteria (PGPR) have emerged as a more sustainable alternative. Streptomyces saraceticus 31 (‘SS31’), a new strain of biocontrol bacteria, was inoculated into rhizosphere soils of ‘Benifuji’ grape to evaluate its impact on grape roots and berries. The results indicated significant improvements in soil fertility, with higher levels of organic matter, phosphorus, potassium, and nitrate nitrogen compared with those of the controls. Moreover, ‘SS31’ application elicited a notable reduction in soil pH levels, along with a substantial augmentation in the enzymatic activities of both phosphatase and invertase. The grapes treated with ‘SS31’ exhibited a notable increase in the number, length, surface area, and volume of fine roots in both 0- to 10-cm and 10- to 20-cm soil profiles. The application of ‘SS31’ resulted in the observation of greater diameter, lower density, and larger lumen area, along with increased specific hydraulic conductivity in the vessels of roots with 1- to 2-mm diameters. Despite a slight reduction in berry weight compared with that of the controls, ‘Benifuji’ grape berries displayed higher total soluble solids and lower total titratable acidity after ‘SS31’ application. Furthermore, ‘SS31’ treatment elevated the levels of volatile compounds in berries, especially fatty acid-derived compounds. A network analysis revealed a robust positive correlation between the observed improvements in grape berry quality and the morphology as well as the hydraulic conductivity of the grape fine roots. In conclusion, these findings suggest that ‘SS31’ has the potential to enhance grape root function by expanding the root absorption area and facilitating water transportation. This, in turn, may improve the flavor and aroma of ‘Benifuji’ grape berries.