Three chloroplast DNA (cpDNA) sequences [the rpl16 intron, the trnL-F, and atpB-rbcL intergenic spacer (IGS)] were employed to study phylogenetic relationships in the genus Rosa. Phylogenetic analyses using these three concatenated sequences were performed using maximum parsimony (MP) and Bayesian inference (BI) methods. Both analyses results suggest that the molecular phylogeny conforms closely to the conventional classification of botanical sections. Morphological similarities between R. sects. Synstylae and Chinenses, and R. sects. Rosa and Carolinae are corroborated on the molecular level in our analyses. Four taxa from R. sect. Pimpinellifoliae are further divided into two small clades, which reflect the morphological characters for these species on a molecular level. Whereas three accessions of R. foetida from R. sect. Pimpinellifoliae form a separate clade. R. ×fortuniana forms a clade with R. laevigata based on its maternal inheritance of cpDNA. R. ×cooperii is hypothesized to be a hybrid with seed parent from R. sects. Synstylae and Chinenses. And R. roxburghii should be classified as a section within the R. subgen. Rosa, rather than being treated as its own subgenus, based on molecular analyses.
Chengyuan Liu, Guoliang Wang, Hui Wang, Tao Xia, Shouzhou Zhang, Qigang Wang, and Yanming Fang
Weibing Zhuang, Xiaochun Shu, Xinya Lu, Tao Wang, Fengjiao Zhang, Ning Wang, and Zhong Wang
Tao Wang, Ruijie Hao, Huitang Pan, Tangren Cheng, and Qixiang Zhang
Mei (Prunus mume) is widely cultivated in eastern Asia owing to its favored ornamental characteristics and its tolerance for low temperatures. Reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR) is a widely used method for gene expression analysis, requiring carefully selected reference genes to ensure data reliability. The aim of this study was to identify and evaluate reference genes for qRT-PCR in mei. Ten candidate reference genes were chosen, and their expression levels were assessed by qRT-PCR in four sample sets: 1) flowering mei; 2) mei undergoing abiotic stress; 3) different genotypes of Prunus species; and 4) all mei samples. The stability and suitability of the candidate reference genes were validated using commercially available software. We found that protein phosphatase 2A-1 (PP2A-1) and PP2A-2 were suitable reference genes for flowering with ubiquitin-conjugating enzyme E2 (UBC) also being suitable for different genotypes of Prunus species. UBC and actin (ACT) were most stably expressed under abiotic stress. Finally, the expression of an AGAMOUS homolog of Arabidopsis thaliana (PmAG) and a putative homolog of Group 2 late embryogenesis abundant protein gene in A. thaliana (PmLEA) were assessed to allow comparisons between selected candidate reference genes, highlighting the importance of careful reference gene selection.
Yalong Qin, Yiming Chen, Weibing Zhuang, Xiaochun Shu, Fengjiao Zhang, Tao Wang, Hui Xu, Bofeng Zhu, and Zhong Wang
Jinxin Wang, Tao Luo, He Zhang, Jianzhu Shao, Jianying Peng, and Jianshe Sun
Hormones have an important role in apple flower bud differentiation; therefore, it is necessary to systematically explore the dynamic changes of endogenous hormones during flower and leaf bud development to elucidate the potential hormone regulation mechanism. In this study, we first observed the buds of ‘Tianhong 2’ apple during their differentiation stage using an anatomical method and divided them into physiologically differentiated stages of spur terminal buds, flower buds, and leaf buds. Then, we determined the contents of zeatin riboside (ZR), abscisic acid (ABA), auxin (IAA), and gibberellin (GA3) in these various types of buds using an enzyme-linked immunosorbent assay. The results showed that the content of ZR and the ratio of ZR to IAA in spur terminal buds decreased significantly during physiological differentiation. The contents of ZR, IAA, and GA3 in leaf buds culminated at the initial differentiation stage. The content of ZR in flower buds was significantly higher than that in leaf buds after formation of the inflorescence primordium and sepal primordium. Before the appearance of stamen primordium, the content of GA3 in flower buds was remarkably lower than that in leaf buds. The ratios of ABA/IAA and ZR/IAA in flower buds were significantly higher than those in leaf buds before the appearance of flower organ primordium. Moreover, ABA content, ABA/ZR, and ABA/GA3 in flower buds were higher than those in leaf buds throughout the whole flower bud morphological differentiation process. Therefore, the reduced ZR content was beneficial to floral induction. The low content of GA3, and high ratios of ABA/IAA and ZR/IAA were conducive to early morphological differentiation. In addition, high ratios of ABA/GA3 and ABA/ZR were beneficial to the morphological differentiation of flower buds. Moreover, the high ABA content was beneficial to floral induction and morphological differentiation of flower buds. Our results shed light on the mechanisms of hormonal regulation of apple flower bud differentiation and could potentially strengthen the theoretical basis for artificial regulation of apple flower bud development using exogenous plant hormones.
Shutian Tao, Danyang Wang, Cong Jin, Wei Sun, Xing Liu, Shaoling Zhang, Fuyong Gao, and Shahrokh Khanizadeh
Lignin is the main component of stone cells, and stone cell content is one of the crucial factors for fruit quality in chinese white pear (Pyrus ×bretschneideri). The lignin biosynthesis pathway is complex and involves many enzymatic reactions. Cinnamate-4-hydroxylase [C4H (EC.188.8.131.52)] is an essential enzyme in lignin metabolism. This study was conducted to investigate the effect of bagging on lignin metabolism during fruit development in chinese white pear. The study showed that bagging had little effect on stone cell content, lignin content, C4H activity, and C4H gene expression and that there was a positive correlation between C4H gene expression and lignin content as well as stone cell content. Moreover, a full-length complementary DNA (cDNA) encoding C4H (PbrC4H, GenBank accession number KJ577541.1) was isolated from chinese white pear fruit. The cDNA is 1515 bp long and encodes a protein of 504 amino acids. Sequence alignment suggested that the deduced protein belongs to the P450 gene family and that C4H might be located subcellularly in the cell membrane. The results indicate that bagging cannot change the lignin and stone cell content significantly and that C4H catalyzes a step in lignin biosynthesis. These findings provide certain theoretical references and practical criteria for improving the quality of chinese white pear.
Dao-Jing Wang, Jing-Wen Zeng, Wen-Tao Ma, Min Lu, and Hua-Ming An
Rosa roxburghii Tratt (Rosaceae) of various organ surfaces are widely existing trichomes. Certain varieties have fruits that are thickly covered with macroscopic trichomes. R. roxburghii Tratt (RR) and R. roxburghii Tratt. f. esetosa Ku (RRE) are important commercial horticultural crops in China because of their nutritional and medicinal values. RRE is generally considered a smooth-fruit variant that arose from RR. Despite their economic importance, the morphological and anatomic features of organ trichomes have not been explored in detail for these two rose germplasms. In this research, we investigated the distribution, morphology, and structure of trichomes distributed on the stem, pedicel, fruit, sepal, and marginal lobule sepals (MLS) of RR as well as RRE. This was accomplished using scanning electron microscopy (SEM). There are various shapes of trichomes distributed on the surfaces of stems, pedicels, fruits, and sepals of the two germplasms. Binate prickles arose on the stem nodes in both germplasms, but acicular trichomes, papillary trichomes, and ribbon trichomes were present only on the surfaces of pedicels in RR. Likewise, flagelliform trichomes were present only on the surfaces of pedicels in RRE. Furthermore, a transection of stems shows that thorns in the two germplasms are composed of epidermis, meristematic layer, and parenchyma cells. The trichome epidermis and meristematic layer in stems of RR are composed of round cells, whereas RRE exhibits square cells in the same layers. Trichomes on the fruit of RR were macroscopic and of single flagelliform and acicular shape. RRE exhibited polymorphic trichomes of flagelliform, triangular, capitate glandular, and elliptic glandular shapes on the pericarp. On the surfaces of RR sepals, there are thick macroscopic acicular trichomes. In contrast, RRE sepals presented flagelliform trichomes and capitate glandular trichomes. It is interesting that no trichomes were found on the surfaces of the MLS in the two germplasms; however, stomata were densely packed on the MLS of RRE when compared with RR. For RR, the trichomes on both sepal and fruit are composed of an epidermis layer and parenchyma cells; however, the epidermis cells of sepal trichomes are polygon-shaped, in contrast to the round epidermis cells in fruit. These results suggest that the two rose germplasms are good candidates for understanding the trichome ontogeny in the genus and for further breeding of the smooth organ trait in this rose species.
Yingli Ma, Tingting Yuan, Tao Wang, Jiaxin Li, Zhongqiu Xu, Siqian Luo, and Yinfeng Xie
In the actual cultivation process, blind fertilizer application was widespread, resulting in a serious decline in the yield of Pseudostellaria heterophylla. We used the 3414 fertilizer experiment design to study the effects of combined Boron (B), Molybdenum (Mo), and Copper (Cu) on the growth indexes, diurnal changes of photosynthesis, and rapid fluorescence induction dynamics in P. heterophylla. Our results show that the optimal combination of B, Mo, and Cu simultaneously promoted the growth of underground and aboveground parts, and significantly improved the quality of single root tuber and yield per unit area. The best combination was treatment 9 (T9 = B, 1 g/L; Mo, 0.08 g/L; Cu, 0.05 g/L), and resulted in a 35.1% increase in yield per unit area compared with the control group (T1). Although the optimal combined application of microfertilizers did not change the bimodal trend of diurnal variation of photosynthesis, it effectively increased the daily average, peak, and valley values of the photosynthetic rate by alleviating the nonstomatal limitation and the photosynthetic midday depression. Pseudostellaria heterophylla leaves showed greater photochemical activity and less photoinhibition of photosystem II in T9. Major effects were that it helped protect the activity of the oxygen-evolving complex to reduce the oxidative damage of chloroplasts and prevent the dissociation of thylakoid. The microfertilizer application also enhanced the electron receiving ability of the QB and plastoquinone (PQ) electronic pools, thereby increasing the ability of electron transfer from QA to QB. The number of reaction centers per unit area was promoted notably by the fertilization treatment.
Raymond Fung*, Chien Wang, David Smith, Kenneth Gross, Yang Tao, and Meisheng Tian
Methyl salicylate (MeSA) and Methyl jasmonate (MeJA) treatments increased chilling resistance of light red tomato (Lycopersicon esculentum cv. Beefsteak) and extended shelf life and fresh-cut quality. We previously showed induction of AOX expression by low temperature and that induction of AOX transcript by MeSA and MeJA is correlated with resistance against chilling injury in peppers. Here, we investigate tomato, which is genetically closely related to peppers and belongs to the same Solanaceae family. In particular, we used four EST tomato clones of AOX from the public database that belong to two distinctly related families, 1 and 2 defined in plants. Three clones designated as LeAOX1a, 1b and 1c and the fourth clone as LeAOX2. Probes for these four genes were designed and Southern blotting done to confirm that they do not cross-hybridize. We will present data from Southern, Northern hybridization and RT-PCR to show: (1) gene copy number of each of these AOX members in the tomato genome; (2) gene-specific expression profiles in response to MeSA and MeJA in cold stored tomato; and (3) the relative transcript abundance of these four AOX genes.
Zhuang-Zhuang Liu, Tao Chen, Fang-Ren Peng, You-Wang Liang, Peng-Peng Tan, Zheng-Hai Mo, Fan Cao, Yang-Juan Shang, Rui Zhang, and Yong-Rong Li
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.