`Pingyitiancha' has good agricultural traits and is an important rootstock for apples. It was studied for Kam, Cef and Carb concentrations, precultural times, active medium of A.tum, Ca++ concentration, mixed inoculation times of AgNO3, and PVP delay selection times for getting transgenic `Pingyitiancha' stock with more resistance to drought and salt. It was demonstrated that the highest transformation frequency was obtained with 200 mg·L-1 Cef as an A.tum restraining antibiotic, 8 mg·L-1 Kam as a selection antibiotic, GCJ8 active medium, a precultural of 2 days, 5 Ca++ concentrations in preculture and coculture medium compared with standard MS medium, immersed in A. tum as OD600 for 0.5 to 1 min., 4 mg·L-1 AgNO3 and 0.7% PVP as an anti-oxidation compound to reduce hydroxybenzene oxidation and delay selection for 5 days by using `Pingyitiancha' apex shoots as explants. Using mediate of A. tum strain LBA4404 under conditions mentioned above and same explants, the HVA1 gene were transformed into `Pingyitiancha' under the control of CaMV35S promoter and obtained Kam resistance regeneration plants. Six transgenic plants were confirmed by PCR and subsequent dot blot methods.
Xiang Shen*, Zhenlin Wei, and Ling Guo
Lulu Zhang, Yunfei Mao, Yunyun Wang, Lu Yang, Yijun Yin, Xiang Shen, Canhong Zhang, and Duojiao Zhang
Lulu Zhang, Yijun Yin, Yunfei Mao, Yeping Liu, Huiling Pang, Xiafei Su, Yanli Hu, and Xiang Shen
Haiyan Wang, Rong Zhang, Weitao Jiang, Yunfei Mao, Xuesen Chen, Xiang Shen, Chengmiao Yin, and Zhiquan Mao
The study here aimed to investigate the effects of pre-winter ditching and freezing-thawing on soil microbial structure in different soil layers of old apple orchards. A total of 30 samples were obtained from 3 Nov. 2016 to 9 Mar. 2017. The relative abundance, alpha diversity, community structure of fungi, and the relationship between environmental factors and microbial community structure were analyzed, and the greenhouse experiments were used for further verification. Results showed that the number of actinomycete and total bacterial colonies decreased, whereas the number of fungi sustained decreased, resulting in a higher bacteria/fungi ratio. The percentage of Fusarium initially decreased, then later increased by 11.38%, 3.469%, 2.35%, 2.29%, and 3.09%. However, Fusarium levels were still 9% lower on 9 Mar. 2017 that on 3 Nov. 2016. Both the abundance and diversity of the community were higher in the upper soil than in the lower. The main environmental factor contributing to the percentage of Fusarium change was average temperature (AT), although highest temperature (HT) and water content (WC) also had an impact. The Malus hupehensis Rehd. seedlings growing in lower soil were more vigorous than that in upper soil. In sum, pre-winter ditching and freezing-thawing in old apple orchards can reduce the abundance percentage of harmful Fusarium and promote the growth of M. hupehensis Rehd. seedlings.
Yijun Yin, Xueli Cui, Lulu Zhang, Yunfei Mao, Xiafei Su, YePing Liu, Huiling Pang, and Xiang Shen
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.
Ran Chen, Weitao Jiang, Haiyan Wang, Fengbing Pan, Hai Fan, Xuesen Chen, Xiang Shen, Chengmiao Yin, and Zhiquan Mao
Apple replant disease (ARD) has been reported in all major fruit-growing regions of the world and is often caused by biotic factors (pathogen fungi) and abiotic factors (phenolic compounds). Soil chemical fumigation can kill soil pathogenic fungi; however, the traditionally used fumigant methyl bromide has been banned because of its ozone-depleting effects. There is thus a need to identify greener fumigant candidates. We characterized the effects of different fumigants on the replanted soil environment and the growth characteristics of Malus hupehensis Rehd. seedlings. All five experimental treatments [treatment 1 (T1), metham-sodium; treatment 2 (T2), dazomet; treatment 3 (T3), calcium cyanamide; treatment 4 (T4), 1,3-dichloropropene; and treatment 5 (T5), methyl bromide] promoted significantly the biomass, root growth, and root respiration rate of M. hupehensis seedlings and the ammonium nitrogen (NH4 +-N) and nitrate nitrogen (NO3 –-N) contents of replanted soil. Metham sodium (T1) and dazomet (T2) had stronger effects compared with 1,3-dichloropropene (T4) and calcium cyanamide (T3). At 172 days after T1, the height, root length, and root respiration rate of Malus hupehensis Rehd. seedlings, and the NH4 +-N and NO3 –-N contents of replanted soil increased by 91.64%, 97.67%, 69.78%, 81.98%, and 27.44%, respectively, compared with the control. Thus, dazomet and metham sodium were determined to be the optimal fumigants for use in practical applications.
Haiyan Wang, Ran Chen, Yuefan Sheng, Weitao Jiang, Rong Zhang, Xuesen Chen, Xiang Shen, Chengmiao Yin, and Zhiquan Mao
The relationship between soil texture and the degree of apple replant disease (ARD) was analyzed from the perspective of the microbial community structure and diversity within the rhizosphere soil of Malus hupehensis Rehd. seedlings. Three different textured soils were taken from different apple orchards in Laizhou, Yantai. The soils were divided into two parts, one was kept in replanted conditions, and the other was fumigated with methyl bromide to act as a high standard control. The strength of ARD occurrence was examined by measuring fresh and dry weight suppression (%) of the M. hupehensis seedlings. Differences in the fungal community structure (especially in Fusarium) among the three soil texture types were analyzed using high-throughput sequencing. The results showed that replanted loam clay soil had the highest fungal diversity, followed by sandy loam soil and finally loam soil. The richness of fungi between soil textures, however, was not significantly different. At the genus level, the relative abundance of Fusarium was 1.96%, 0.78%, and 10.89% in replanted sandy loam, replanted loam soil, and replanted loam clay soil, respectively. Moreover, the gene copy number of Fusarium oxysporum, Fusarium solani, and the inhibition rate of fresh weight of M. hupehensis seedlings were the same in the three soil textures. The plant height, photosynthesis (net) (Pn), and stomatal conductance (g S) of the M. hupehensis seedlings were significantly less in the replanted soil compared with the control treatments, with the overall difference being greatest in replanted loam clay soil, followed by replanted sandy loam and then replanted loam soil.
Kai-Xiang Li, Kai Liu, Yingying Chen, Xiaolu Huang, Wenhui Liang, Baocai Li, Yingbai Shen, and Haiying Liang
Lithocarpus polystachyus is a unique medicinal tree species that is valued for its abundant flavonoids in leaves. Currently, genes and metabolites involved in the flavonoid biosynthesis pathway remain largely unknown. To elucidate the flavonoid biosynthesis pathways, transcriptome and metabolome analyses of young, mature, and old leaves were conducted. A total of 86,927 unigenes were obtained, and 51.4% of them were annotated in eight public databases. The majority of the 44 candidate genes in the flavonoid biosynthesis pathway were downregulated as leaves aged. Metabolome profiling revealed a set of 427 metabolites in leaves. Consistent with the transcriptome results, 15 of the 19 metabolites in the flavonoid pathway decreased during the development of leaves. The data indicate that young leaf is the optimal stage for tea harvest. This is the first report of integrated transcriptome and metabolome profiling of L. polystachyus. This study demonstrates the correlation of gene expression and metabolites related to flavonoid biosynthesis and reveals the key genes responsible for flavonoid accumulation in young leaf. The information can be applied to future studies performed to elucidate and manipulate flavonoid biosynthesis in L. polystachyus.
Li-Qiang Tan, Xin-Yu Wang, Hui Li, Guan-Qun Liu, Yao Zou, Shen-Xiang Chen, Ping-Wu Li, and Qian Tang
Landrace tea populations are important recourses for germplasm conservation and selection of elite tea clone cultivars. To understand their genetic diversity and use them effectively for breeding, two traditional landrace tea populations, Beichuan Taizicha (BCTZ) and Nanjiang Dayecha (NJDY), localized to northern Sichuan, were evaluated for morphological characters, simple sequence repeat (SSR)–based DNA markers and the contents of biochemical components. A wide range of morphological variation and a moderately high level of DNA polymorphism were observed from both BCTZ and NJDY. NJDY had on average, bigger leaves, larger flowers, higher total catechins (TCs), and greater gene diversity (GD) than BCTZ. Interestingly, samples from BCTZ had a wide range in the ratio of galloylated catechins to nongalloylated catechins (G/NG) (1.83–8.12, cv = 48.8%), whereas samples from NJDY were more variable in total amino acid (TAA) content (25.3–50.8 mg·g−1 dry weight) than those from BCTZ. We concluded that the two Camellia sinensis landrace populations are of great interest for both individual selection breeding and scientific studies.