MYB (v-myb avian myeloblastosis viral oncogene homologs) transcription factors (TFs) are involved in diverse physiological processes, including cell shape determination, cell differentiation, and secondary metabolism, as well as abiotic stress response. In the present study, MdMYB4, an R2R3-MYB protein that is a homolog of Arabidopsis thaliana MYB4, was identified and characterized. Quantitative real-time polymerase chain reaction (qRT-PCR) expression analysis demonstrated that MdMYB4 is extensively expressed in various apple (Malus domestica) tissues and that its expression is induced by cold, osmotic, and salt stress. An MdMYB4-GFP fusion protein was localized in the nucleus of transformed onion (Allium cepa) epidermal cells and had a certain transcriptional activation activity by yeast one-hybrid assay. Overexpression of the MdMYB4 gene remarkably enhanced the tolerance of stably transgenic apple calli to severe salt and cold stress, and both the relative conductivity and malondialdehyde (MDA) accumulation of transgenic calli under salt and cold stress were significantly lower than in the wild type control. Taken together, these results suggest that MdMYB4 may play a positive regulatory role in both cold and salt stress responses.
Ruigang Wu, Yi Wang, Ting Wu, Xuefeng Xu, and Zhenhai Han
Ying Fang, Ting Lei, Yanmei Wu, and Xuehua Jin
The calla lily (Zantedeschia hybrida) is a valued ornamental plant due to its unique shape and color variations. To determine the mechanisms responsible for color development in the calla lily spathe, we conducted a comparative transcriptomic analysis of the spathes of the black [Black Girl (B)], pink [Romantic (P)], and white [Ventura (W)] cultivars. The gene expression patterns in six spathe colors, including the preceding three colors as well as the amaranth [Promise (N)], red [Figo (F)], and yellow [Sun Club (Y)] cultivars were analyzed by real-time quantitative polymerase chain reaction (PCR). Transcriptomic analysis identified 25,165 differentially expressed genes. The transcription abundance and expression level of genes annotated as anthocyanidin reductase (ANR1, ANR2), basic-helix-loop-helix (bHLH1), and glutathione S-transferases (GST1) were significantly upregulated in B, and the expression of anthocyanidin synthase (ANS) was highest in B except for N. However, chalcone isomerase (CHI2) and dihydroflavonol 4-reductase (DFR1, DFR2) were expressed at significantly lower levels in P, W, and Y. Correlation analysis revealed that bHLH1 might act as a positive regulator of ANS expression, promoting anthocyanin synthesis. Moreover, GST1-encoded proteins may be related to the accumulation and transport of both anthocyanin and procyanidin in the calla lily spathe. It is speculated that the formation of the black spathe is related to the accumulation of anthocyanins and procyanidins. However, the low expression of CHI2, DFR1, and DFR2 may result in the inhibition of anthocyanin synthesis, which may lead to lightening of the spathe color. This preliminary study revealed the mechanism responsible for calla lily spathe color, identifying the key genes involved, thus providing effective gene resources and a theoretical basis for flower color molecular breeding.
Yi Tan, Baisha Li, Yi Wang, Ting Wu, Zhenhai Han, and Xinzhong Zhang
Agrobacterium-mediated genetic transformation is commonly used in dicotyledon plants such as apples. The regeneration ability of the recipient is an important factor in transformation efficiency. Here, the variations in bud regeneration rate (BRR) and the number of adventitious buds (NAB) formed per explant in Malus germplasm accessions with phenological stage were estimated. Both BRR and NAB of explants at the dormancy broken and spring sprouting stages were significantly higher than those at the autumn sprouting stage. The genetic diversity and inheritance of BRR and NAB were evaluated using 153 Malus germplasm accessions and 78 hybrid trees of Jonathan × Golden Delicious. Malus sieversii 31, Liberty, and Smoothee exhibited significantly high BRR (98.33%, 98.33%, and 93.33%, respectively) and a large NAB without vitrification. BRR and NAB linearly correlated with each other but not with callus formation rate. The broad sense heritability of the regeneration rate was 92.16%. The three Malus accessions that had high regeneration ability, and some of their sexual descendants, might be outstanding genetic resources for future genetic transformation.
Keng Heng Chang, Rung Yi Wu, Geng Peng Chang, Ting Fang Hsieh, and Ren Shih Chung
Coir is used around the world as a cultivation medium for plants; its commercial popularity is the result of its availability, low cost, and environmentally friendly characteristics. It is used as a medium in the hydroponic cultivation of Anthurium (Anthurium andraeanum Lind.) in Taiwan and is a new source for cut flower production around the world. Little is known about the nutrient requirements of Anthurium cultivated in coir under fluctuating climatic conditions. The objective of this study was to evaluate the influences of various nitrogen (N) concentrations on the growth and nutrient uptake of Anthurium cultivated in coir under different seasonal conditions. Four levels of N concentration in nutrient solution were used: 79 mg·L−1 (NS79 treatment), 105 mg·L−1 (NS105 treatment), 158 mg·L−1 (NS158 treatment), and 210 mg·L−1 (NS210 treatment) with NS105 serving as the control. The effects of N concentration and seasonal fluctuations on Anthurium were measured in dry weight, leaf growth, flower growth, and nutrient uptake at different growth stages during the 2-year study period. The results show that the dry weight, leaf area, and flower number were higher in plants receiving NS105 and NS158 treatments than those receiving NS79 and NS210 treatments. However, the NS158-treated plants produced better quality cut flowers than the NS105-treated plants in the first year of cultivation as indicated by their wider, circular spathe. Retarded growth of NS79-treated Anthurium was the product of insufficient N supply and reduced carbon (C) assimilation. The excess supply of N in the NS210 treatment resulted in small potassium (K) and magnesium (Mg) uptakes, which in turn resulted in poor growth in the second year of cultivation. However, the nutrient supplies in the NS158 and NS210 treatments yielded better Anthurium growth during the initial stage than the NS79- or NS105-treated groups. Regardless of plant growth, flower yield, and nutrient uptake, there were significant interactions between N treatments and seasonal fluctuations in subtropical conditions during year-round cultivation. We concluded that the limiting factor in Anthurium growth and yield during the spring and summer is the N supply, whereas climate conditions are the limiting factor during the fall and winter.
Haishan An, Feixiong Luo, Ting Wu, Yi Wang, Xuefeng Xu, Xinzhong Zhang, and Zhenhai Han
Fine root (≤2 mm in diameter) systems play a pivotal role in water and mineral uptake in higher plants. However, the impact of fine root architecture on tree growth and development is not fully understood, especially in apple trees. Here, we summarize a 6-year-trial study using minirhizotrons to investigate the relationships between fine root production, mortality, and longevity in ‘Red Fuji’ trees grafted on five different rootstocks/interstems. Based on root length density (RLD), fine root production and mortality were markedly lower in ‘Red Fuji’ trees growing on dwarfing M.9 (M.9) and Shao series no. 40 (SH.40) rootstocks than in trees on standard Malus robusta ‘Baleng Crab’ (BC) rootstock. The use of M.9 and SH.40 as interstems led to an extensive reduction in fine root production and mortality in comparison with BC rootstock. Root number density (RND), but not average root length (ARL), showed similar patterns to RLD. About one-half of fine roots in ‘Red Fuji’ tree growing on M.9 were scattered within the top 0–20 cm of topsoil, indicating shallow root system in M.9, whereas in trees on BC, 55.15% of fine roots were distributed between 100- and 150-cm soil depth, indicating a deep root architecture. The addition of interstems did not alter fine root soil-depth distribution. For all rootstocks/interstems, fine roots with a life span of less than 80 days were generated in spring and summer, but fine roots which lived for more than 81 days were produced almost all the year round. In conclusion, lower fine root numbers were associated with the dwarfing effect in dwarfing rootstocks/interstems, but ARL and shallower rooting were not.
Meiling Zhang, Ming Chen, Zhen Wang, Ting Wu, Yi Wang, Xinzhong Zhang, and Zhenhai Han
Grafting has been widely used in orchard management because the rootstock can make the tree more tolerant to environmental stresses. Iron deficiency is one of the major limiting environmental factor in apple production worldwide. Systematic research has been made about iron-deficiency adaptive responses in the level of organs, cells, and subcells, whereas the interactions between Fe and other divalent cations in tissue level are little known. Synchrotron radiation X-ray fluorescence (SR-μXRF) was used to map the location of selected elements Fe, Zn, Mn, Ni, and Co in the longitudinal and latitudinal root samples of Malus xiaojinensis. Iron deficiency induced a significant increase in the relative contents of five micronutrients in epidermis and cortex. The ratio of element contents of roots under Fe-deficient condition and Fe-sufficient condition at same position increased obviously in the section of 1000- to 2000-μm distance from the root tip in xylem. Expression analysis of iron absorption- and transport-related genes in roots showed that MdNramp3 and MxCS1 increased significantly. These results indicated that iron deficiency promoted the long-distance transport of micronutrients in xylem, and MdNramp3 and MxCS1 might play an important role in this process. Importantly, this study directly provides visual divalent metals distribution in tissue level for an improved understanding of metal absorption process in apple rootstock.
Chenping Zhou, Ruiting Chen, Yaqiang Sun, He Wang, Yi Wang, Ting Wu, Xinzhong Zhang, Xuefeng Xu, and Zhenhai Han
Bridge grafting is widely applied in trunk-wounded apple trees. In this study, we carried out semigirdling and ring girdling on the trunk of ‘Nagafu 2’/Malus baccata (L.) Borkh apple trees to simulate trunk injury. We then bridge grafted a M9 self-rooted rootstock on the injured trunks to study the effects of bridge grafting on flowering, fruit-set, tree vigor, and fruit characteristics in ‘Nagafu 2’ apple. The results showed that both semigirdling and ring girdling due to the large wounded area caused significant decrease in flowering, fruit-set, and tree vigor (estimated by measuring leaf area, leaf gas exchange, tree height, and shoot growth); in addition, ring girdling increased flesh and peel firmness. However, bridge grafting of M9 self-rooted rootstock on semigirdling and girdling apple trees resulted in partial recovery of tree vigor (leaf area and photosynthesis) and maintaining the reduction of vegetative growth, thereby increasing flowering, fruit-set, yield, fruit weight, and peel firmness.