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.
Tao Wang, Ruijie Hao, Huitang Pan, Tangren Cheng and Qixiang Zhang
Ying Kong, Ming Sun, Hui-tang Pan and Qi-xiang Zhang
Floral scents emitted from eight cultivars of cut lily flowers (Lilium) were analyzed. Floral volatiles were collected by headspace adsorption on sorbent tubes and analyzed by gas chromatography–mass spectrometry (GC/MS) using a direct thermal desorption. Fifty volatile compounds were identified. Nine compounds were detected in all lilies, whereas 20 compounds were detected in all scented lilies. The results revealed that non-scented lilies emitted trace amounts of volatile compounds, whereas scented lilies emitted high levels of volatile compounds. Monoterpenoids and benzenoids were the dominant compound classes of volatiles emitted from scented lilies. Myrcene, (E)-β-ocimene, linalool, methyl benzoate, and ethyl benzoate were the major compounds of the aroma of scented lilies; 1,8-cineole was also a major compound in the two scented oriental × trumpet hybrid lilies. Scent emissions occurred in a circadian rhythm with higher levels of volatiles emitted during the night. Lilium ‘Siberia’ was selected as a model to investigate the source of the emissions. GC/MS analysis of four flower parts and neutral red staining revealed that tepals were the source of floral scent.
Le Luo, Chao Yu, Xuelian Guo, Huitang Pan and Qixiang Zhang
Rosa laxa is widely distributed in the Xinjiang Uygur Autonomous Region of China and is highly adaptable and rich in variation. In this study, we investigated the morphology, habitats, and palynomorphology of R. laxa botanical varieties from Xinjiang, China. In addition to R. laxa var. laxa, there were three other botanical varieties of R. laxa growing in southern Xinjiang, including var. mollis, var. kaschgarica, and var. tomurensis. Together, these four botanical varieties exhibited morphological variation, mainly in the morphology of prickles and the glandular trichome and in flower color. The pollen grains of the R. laxa botanical varieties, all medium in size (21.77–48.39 μm), came in three shapes: perprolate, prolate, and subspheroidal. Their pollen exine sculptures were characterized by either a striate-perforation pattern or striate pattern, but perforation varied in terms of diameter and density and striae varied in depth and density. Palynomorphological assessment showed that three types of evolution, i.e., primitive, transitive, and evolved, were present among R. laxa botanical varieties, and pollen dimorphism was observed in the same botanical variety. Perprolate pollen with a dense striate pattern was the most evolved type. Based on morphological and palynomorphological investigations, var. tomurensis was considered to be the most evolved one among the studied botanical varieties.
Ming Cai, Ke Wang, Le Luo, Hui-tang Pan, Qi-xiang Zhang and Yu-yong Yang
Hydrangea macrophylla is the most popular species in the genus Hydrangea because of its large and brightly colored inflorescences. Since the early 1900s, numerous cultivars with showy flowers have been selected. Although many H. macrophylla cultivars have been developed, cold hardiness is still the major limitation to their outdoor use. Hydrangea arborescens is a small attractive shrub or subshrub native to northeastern parts of the United States, which is valued for its hardiness. Interspecific breeding of H. arborescens and H. macrophylla has been tried, but putative hybrid seedlings either died at an early stage or were not verified. We made successful hybridizations between H. macrophylla ‘Blue Diamond’ and H. arborescens ‘Annabelle’ and used in vitro ovary culture to produce viable plants. Hybrids were intermediate in appearance between parents, but variable in leaves, inflorescences, and flower color. The success of this hybridization was confirmed by six simple sequence repeat (SSR) genetic markers. The maternal chromosome number was 36, and the paternal number was 38. Chromosome counts of hybrids indicated that nearly half of them were aneuploids. Male fertility of progeny was evaluated by fluorescein diacetate staining of pollen. Twelve out of 14 hybrids (85.7%) had male fertility. We documented the first flowering progeny of H. macrophylla and H. arborescens, suggesting an effective beginning to a cold hardiness breeding program.
Le Luo, Yichi Zhang, Yingnan Wang, Tangren Cheng, Huitang Pan, Jia Wang and Qixiang Zhang
Gesnariad (Primulina yungfuensis) is a popular houseplant species, native to southwest China. However, stunting frequently occurs as a result of limited knowledge about the growth requirements of this plant. Understanding water and fertilizer requirements of gesnariad are important for effective large-scale greenhouse cultivation. Using a response surface methodology (RSM) based on a rotatable central composite design (RCCD; half implementation), a pot experiment was performed in a natural-light greenhouse from June to Sept. 2014. The study assessed the interaction between irrigation volume (W) and nitrogen (N), phosphorus (P), and potassium (K) fertilizer rates on plant height, crown diameter, number of leaves, single leaf area, and fresh weight. Results showed that W had a significant positive effect on plant height, crown diameter, single leaf area, and fresh weight. Furthermore, P fertilization resulted in increased leaf number. Combined P and K fertilization reduced individual leaf area, whereas combined N and P fertilization reduced fresh weight. By analyzing the multiobjective decision-making model, we found that a combination of 100.2 mL water, 3.6 mmol·L−1 N, 0.10 mmol·L−1 P, and 1.2 mmol·L−1 K could be used to achieve optimum growth of gesnariad.
Cunquan Yuan, Zhiyi Qu, Huitang Pan, Tangren Cheng, Jia Wang and Qixiang Zhang
Heterostylous Primula forbesii is an important ornamental flower in China because of its long-lasting flowers and winter bloom. This study aimed to develop markers of expressed sequence tag–simple sequence repeats (EST-SSRs) that are associated with heterostyly and that can be used for molecular-assisted selective breeding in P. forbesii. We investigated 114,474 unigenes and identified 25,095 SSRs in P. forbesii. Dinucleotide repeats (46.14%), mononucleotide repeats (44.65%), and trinucleotide repeats (8.27%) were the most abundant SSRs. Among the 25,095 SSRs, 10,645 SSR primer pairs were successfully designed, of which 130 primer pairs were randomly selected for further amplification validation using eight accessions of P. forbesii; 98 pairs produced clear and stable polymerase chain reaction (PCR) products, and 28 pairs showed polymorphism. Bulked segregant analysis (BSA) was conducted for the F1 population with respect to thrum style and pin style by scanning 28 polymorphic SSR primer combinations. One SSR marker, c64326, linked to the heterostyly trait at a genetic distance of ≈3.70 cM was identified. The marker c64326 was further validated in two populations with an accuracy of 97.92% and 90.63%. The novel and linked EST-SSR markers can be valuable resources for genetic diversity analysis, mapping, and marker-assisted breeding in P. forbesii.
Jia-yi Wang, Jian-shuang Shen, Mengmeng Gu, Jia Wang, Tang-ren Cheng, Hui-tang Pan and Qi-xiang Zhang
Yellow-leafed cultivars usually do not grow as vigorous as their green-leafed counterparts, which affect their use in landscapes. To breed Forsythia cultivars with both yellow leaves and vigorous growth, crosses between F. ‘Courtaneur’ (♀) and Forsythia koreana ‘Suwon Gold’ (♂) were conducted, and 52 F1 hybrid progenies with different leaf colors (green, chartreuse, and yellow) were obtained. The progenies were categorized into three groups [Yellow Group (YG), Chartreuse Group (CG), and Green Group (GG)] based on leaf colors. The growth index (GI) and the number of branches and leaves of YG progenies were significantly lower at 2%, 35%, and 34% of GG progenies. As the leaves changed from green to chartreuse and to yellow, chlorophyll content, leaf thickness, and chlorophyll fluorescence parameters decreased and the chloroplast structures were disintegrated gradually, which influenced the leaf photosynthetic activity and led to weak growth. Compared with yellow-leafed progenies, the leaf chlorophyll content and leaf thickness of chartreuse-leafed progenies were significantly higher at 71% and 9%. The chloroplast structure of stroma lamella of chartreuse-leafed progenies was relatively intact. Carboxylation efficiency (CE), photochemical efficiency of PS II (F v/F m), and the number of branches and leaves of GG progenies were significantly higher than YG progenies; however, they have no significant difference with CG progenies. The results were promising for breeding new forsythia cultivars from moderate growth and chartreuse leaves.
Chao Yu, Le Luo, Hui-tang Pan, Yun-ji Sui, Run-hua Guo, Jin-yao Wang and Qi-xiang Zhang
Wild Rosa species, which are highly variable and locally adapted, are widely distributed in the Xinjiang Uygur autonomous region of China. These species possess many important horticultural traits that are not found in the gene pool of modern cultivated roses. However, little is known about their cytological characteristics, because few of them have been karyologically analyzed. Karyological data of 13 samples of seven wild Rosa taxa, including R. berberifolia, two botanical varieties of R. spinosissima, R. platyacantha, R. beggeriana, R. acicularis, and R. laxa, were investigated by means of squashes of shoot tips. The results showed that seven samples were diploid (2n = 2x = 14), whereas the other six samples were tetraploid (2n = 4x = 28). The karyotypes of all the test samples were composed of m and sm chromosomes, which include 1A, 2A, 1B, and 2B. There were differences in asymmetry index, centromere index, and distribution of relative lengths. The karyotype of the tetraploid R. laxa var. laxa sample from Aksu easily distinguished from the other R. laxa var. laxa samples tested in having the highest asymmetry index and the most evolved karyotype. This Aksu population merits recognition as a new botanical variety of R. laxa. The karyological data, most of which are first reports for their taxa, provide a comprehensive cytogenetic resource that can be used to better understand the taxonomy, evolution, and speciation in the genus Rosa and to identify candidate species for breeding programs.