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Many reports indicate that an abundance of really interesting new gene (RING) play key roles in regulating defense responses against abiotic and biotic stresses in plants. In this study, the cloning and functional characterization of a RING gene, MaRING2, in banana (Musa acuminata) fruit are reported. MaRING2 belongs to the NEP1-interacting protein (NIP) RING-H2 finger protein family. Gene expression profiles revealed that MaRING2 was cold responsive and induced by abscisic acid (ABA) treatment during cold storage. In this study, the MaRING2 under control of the Cauliflower mosaic virus 35S (CaMV 35S) promoter was transformed to tobacco (Nicotiana benthamiana) using agrobacterium (Agrobacterium tumefaciens)-mediated transformation. The resultant MaRING2-overexpressing transgenic plants (35S:MaRING2) exhibited significantly increased tolerance to low temperatures and were hypersensitive to exogenous ABA in terms of germination and early seedling growth. In addition, overexpression of MaRING2 enhanced the expression of stress-responsive genes under normal (before cold stress) or cold conditions. These results demonstrate the biological role of MaRING2 in conferring cold tolerance. Taken together, these results suggest that MaRING2, a C3H2C3-type RING protein, is a positive regulator of the ABA-dependent stress response.
Ferric chelate reductase (FRO) is a critical enzyme for iron absorption in strategy I plants, reducing Fe3+ to Fe2+. To identify FRO family genes in the local Citrus junos cultivar Ziyang Xiangcheng and to reveal their expression model, the citrus (Citrus sp.) genome was searched for homologies of the published sequence CjFRO1. Five FROs were found, including CjFRO1; these were named CjFRO2, CjFRO3, CjFRO4, and CjFRO5, respectively, and cloned via reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequences of five CjFROs contained flavin adenine dinucleotide (FAD)-binding motifs, nicotinamide adenine dinucleotide (NAD)-binding motifs, and 6–10 transmembrane domains, with isoelectric points between 6.73 and 9.46, and molecular weights between 67.2 and 79.9 kD. CjFRO1 and CjFRO2 were predominantly found in the aboveground parts of C. junos, with CjFRO1 highly expressed in leaves, and CjFRO2 largely expressed in stems and leaves. CjFRO3 was less expressed in roots, stems, and leaves. CjFRO4 and CjFRO5 were predominately found in roots. Under iron-deficient conditions, CjFRO4 was significantly and specifically increased in the roots of C. junos, whereas CjFRO1 was upregulated in the roots and leaves.
As a wild apple species native to central Asia, Malus sieversii (Ledeb.) Roem. is distributed in a wide region covering most of the Tienshan Mountains. Malus sieversii is a useful genetic pool for apple breeding since rich with diversity. In this paper, we first describe the species range of this endangered species. We then describe an in situ reserve that has been established. We also investigated some reproductive characteristics of M. sieversii including pollen germination, seed dormancy, and seed viability. Both stratification and seedcoat removal efficiently released seed dormancy and accelerated seed germination. Pollen germination rate is around 60%. Our data suggest that injurious insects and human activities, rather than reproductive characters, limit the renewal of M. sieversii.
Three kinds of expression vectors of a pollen-S determinant were constructed to provide a reference for molecular breeding of self-compatible (SC) Prunus species. An S-haplotype-specific F-box (SFB) protein gene from the ‘Xiaobaixing’ apricot (Prunus armeniaca) was cloned by reverse transcription polymerase chain reaction (RT-PCR) and 3′-rapid-amplification of cDNA ends (3′-RACE). A 1136-bp sequence complementary to the 3′-end of the cDNA (GenBank accession number KP938528.2) with a 912-bp complete open reading frame (ORF) was obtained. The deduced amino acid sequence contained an F-box domain, two variable regions, and two hypervariable regions with structural characteristics similar to SFB in other Rosaceae plants. Sense, antisense, and RNA interference (RNAi) vectors for SFB were constructed by enzyme restriction. The target fragment was restricted using the corresponding restriction enzyme and then directionally inserted between the 35S cauliflower mosaic virus promoter and the nopaline synthase terminator (NOS-ter) of the expression vector pCAMBIA-35S-MCS-NOS-NPTII. The intron-containing hairpin RNA (ihpRNA) was obtained by fusion PCR. The constructed vectors were transferred into Agrobacterium tumefaciens strain LBA4404 by freezing/thawing. The RNAi vector of SFB was also transformed in tobacco (Nicotiana tabacum). The successful construction of these three expression vectors provides a basis for transforming ‘Xiaobaixing’ apricot and the breeding of SC Prunus cultivars.
The characterization of aroma of the 14 main apricot (Prunus armeniaca L.) cultivars in Xinjiang was evaluated using high-performance solid-phase microextraction (HP-SPME) with gas chromatography-mass spectroscopy (GC-MS). A total of 208 volatiles that include 80 esters, 25 aldehydes, 15 terpenes, 21 ketones, 39 alcohols, 27 olefins, and 1 acid were identified from these cultivars. The compounds propyl acetate, 3-methyl-1-butanol acetate, (Z)-3-hexen-1-ol acetate, d-limonene, β-linalool, hexanal, hexyl acetate, butyl acetate, β-myrcene, ethyl butanoate, and β-cis-ocimene were the major compounds responsible for aroma in these cultivars. GC-MS results showed that Kuchexiaobaixing, Guoxiyuluke, and seven other cultivars were characterized by a high level of esters and were considered to be fruity apricot aroma. ‘Luotuohuang’ and ‘Heiyexing’ accumulate high levels of terpenes and exhibited an outstanding floral aroma. Higher levels of alcohols and aldehydes were observed in ‘Danxing’, ‘Sumaiti’, and ‘Kumaiti’. The latter are considered green aroma cultivars. These three types of cultivars with different aroma characteristics can be significantly differentiated by using the principal component analysis (PCA) method. The contributions of volatiles to the apricot aroma were assessed by using the partial least squares regression (PLSR) model. Esters, terpenes, and C6 components were shown to be responsible for the fruity, floral, and green character of fresh apricots, respectively.
The DNA binding with one finger (Dof), as an important transcription factor, plays an important role in growth and development, primary and secondary metabolism, stress resistance, and plant hormone signal transduction. However, the identification and analysis of the Dof transcription factor family in Rosa is rarely reported. In this study, 28 Rosa chinensis Dof (RcDof) members were identified, which were located on seven chromosomes. The RcDofs were divided into 12 subfamilies according to evolutionary analysis. Through motif, gene structure, and cis-acting element analyses of the 12 subfamilies, the functions of RcDofs were analyzed and predicted. Furthermore, the Dof members in R. chinensis ‘Old Blush’ and another three species (Arabidopsis thaliana, Oryza sativa, and Zea mays) were systematically analyzed. Twelve subfamilies were found in these four species and the motifs and gene structures of Dof members in each subfamily were similar, which further proves that the RcDofs analysis is accurate. Through an intra- and interspecies collinearity analysis, it was found that the collinearity between A. thaliana and R. chinensis is closer in comparison. Tissue expression analysis of RcDofs was by quantitative reverse-transcription polymerase chain reaction (PCR). Quantitative real-time PCR analysis showed expressions of the RcDofs are tissue specific. The RcDofs had higher expression in leaves, roots, and flowers than other tissues. Taken together, this study provides valuable information for future research on functional exploration of RcDof genes and molecular breeding in Rosa.
To select resistant germplasm resources and understand the growth and physiological responses of kiwifruit (Actinidia sp.) to drought stress, five species, Actinidia macrosperma (Acma), Actinidia longicarpa (Aclo), Actinidia deliciosa (Acde), Actinidia hemsleyana (Ache), and Actinidia valvata (Acva), were assessed under tissue culture conditions. Rootless seedlings of five species were cultured in a medium containing polyethylene glycol [PEG (formula weight 8000)] to induce drought stress (0%, 5%, 10%, 15%, and 20%). After a 30-day culture, three growth indices [fresh weight (FW), plant height (PLH), and leaf number (LN)] and six physiological indices were determined, and the drought damage index (DDI) was determined. The DDIs of five species increased, and three growth indices decreased with increasing PEG concentrations. The following changes were observed under 20% PEG treatment conditions: superoxide dismutase (SOD) activities increased significantly in Acma, Aclo, and Ache specimens; peroxidase (POX) activities remained stable in Acde, Ache, and Acva specimens; and catalase (CAT) activities increased sharply in Acma and Acva. Furthermore, the results indicated that soluble sugar (SS) content increased slightly in Acma, Aclo, Acde, and Ache but it decreased in Acva specimens. Proline (PRO) content increased significantly in Acma and Acva, and malondialdehyde (MDA) contents tended to increase under drought stress in all five species. Principal component analysis (PCA) results indicated that the order of drought tolerance in the five genotypes examined in this study under tissue culture conditions was as follows: Acma > Acva > Acde > Aclo > Ache. Therefore, we concluded that Acma and Acva are more resilient germplasm resources that represent promising kiwifruit-breeding materials. Furthermore, tolerance to drought stress in these species should be further investigated under orchard conditions.
Head splitting resistance (HSR) in cabbage is an important trait closely related to appearance, yield, storability, and mechanical harvestability. In this study, a doubled haploid (DH) population derived from a cross between head splitting-susceptible inbred cabbage line 79-156 and resistant line 96-100 was used to analyze inheritance and detect quantitative trait loci (QTLs) for HSR during 2011–12 in Beijing, China. The analysis was performed using a mixed major gene/polygene inheritance method and QTL mapping. This approach, which uncovered no cytoplasmic effect, indicated that HSR can be attributed to additive-epistatic effects of three major gene pairs combined with those of polygenes. Major gene and polygene heritabilities were estimated to be 88.03% to 88.22% and 5.65% to 7.60%, respectively. Using the DH population, a genetic map was constructed with simple sequence repeat (SSR) markers anchored on nine linkage groups spanning 906.62 cM. Eight QTLs for HSR were located on chromosomes C4, C5, C7, and C9 based on 2 years of phenotypic data using both multiple-QTL mapping and inclusive composite interval mapping. The identified QTLs collectively explained 37.6% to 46.7% of phenotypic variation. Three or four major QTLs (Hsr 4.2, 7.2, 9.3, and/or 9.1) showing a relatively larger effect were robustly detected in different years or with different mapping methods. The HSR trait was shown to have a complex genetic basis. Results from QTL mapping and classical genetic analysis were consistent. Our results provide a foundation for further research on HSR genetic regulation and molecular marker-assisted selection (MAS) for HSR in cabbage.
Cytosine methylation plays important roles in regulating gene expression and modulating agronomic traits. In this study, the fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) technique was used to study variation in cytosine methylation among seven pecan (Carya illinoinensis) cultivars at four developmental stages. In addition, phenotypic variations in the leaves of these seven cultivars were investigated. Using eight primer sets, 22,796 bands and 950 sites were detected in the pecan cultivars at four stages. Variation in cytosine methylation was observed among the pecan cultivars, with total methylation levels ranging from 51.18% to 56.58% and polymorphism rates of 82.29%, 81.73%, 78.64%, and 79.09% being recorded at the four stages. Sufficiently accompanying the polymorphism data, significant differences in phenotypic traits were also observed among the pecan cultivars, suggesting that cytosine methylation may be an important factor underlying phenotypic variation. Hypermethylation was the dominant type of methylation among the four types observed, and full methylation occurred at higher levels than did hemimethylation in the pecan genomes. Cluster analysis and principal coordinate analysis (PCoA) identified Dice coefficients ranging from 0.698 to 0.778, with an average coefficient of 0.735, and the variance contribution rates of the previous three principal coordinates were 19.6%, 19.0%, and 18.2%, respectively. Among the seven pecan cultivars, four groups were clearly classified based on a Dice coefficient of 0.75 and the previous three principal coordinates. Tracing dynamic changes in methylation status across stages revealed that methylation patterns changed at a larger proportion of CCGG sites from the 30% of final fruit-size (30%-FFS) stage to the 70%-FFS stage, with general decreases in the total methylation level, the rate of polymorphism, and specific sites being observed in each cultivar. These results demonstrated that the F-MSAP technique is a powerful tool for quantitatively detecting cytosine methylation in pecan genomes and provide a new perspective for studying many important life processes in pecan.