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The flowering control of Oncidesa Gower Ramsey ‘Honey Angel’ is important and in-demand by the industry. Therefore, an understanding of the development of inflorescence and vegetative shoot from the leaf axils on the current shoot is required. The internode of a young Oncidesa current shoot between the 0th (at the base of the pseudobulb) and 1st (immediately above the pseudobulb) nodes can enlarge to form a pseudobulb, and the axillary bud on the 0th or -1st (immediately below the 0th node) node can differentiate into an inflorescence bud. The axillary buds on the lower nodes (-2nd to -4th nodes) can remain vegetative. In this study, we investigated the growth and anatomical features of axillary buds at various stages during the growth of the current shoot. We sampled the axillary buds on the 0th to -4th nodes from the current shoots when they were 10, 15, 20, 25, and 30 cm in length for sectioning and anatomical observations. Vegetative buds on the -2nd to -4th nodes grew faster and had more nodes than the inflorescence bud when the current shoot grew from 10 to 25 cm. However, when the current shoot elongated from 25 to 30 cm, the length and node number in the inflorescence bud on the 0th node increased and the inflorescence branch primordia were observable. The length and node number of the inflorescence bud became the same as that of the vegetative buds, which had no further growth, whereas the current shoot grew from 25 to 30 cm. The pseudobulb began to emerge from the leaf sheath (unsheathing) when the current shoot had reached 30 cm in length. Therefore, the time when the pseudobulb started to unsheathe from its subtending leaf was critical for the reproductive growth of Oncidesa Gower Ramsey ‘Honey Angel’ when growth acceleration of the inflorescence bud occurred. Evaluating the current shoot length can be a nondestructive method of estimating the developmental stage of the inflorescence bud.

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.

Cytokinins play an important role in regulating plant growth and development. The cytokinin gene, isopentenyl transferase (ipt), was placed under the control of the ACC oxidase promoter from the LEACO1 gene from Lycopersicon esculentum and introduced into Nicotiana tabacum (cv. Havana) and chrysanthemum (Dendranthema × grandiflorum `Iridon'). Transformants were confirmed by PCR reaction and Southern blot and analyzed for phenotypical changes under both greenhouse and growth chamber conditions. With both species, LEACO1-ipt transgenic plants displayed a wide range of vegetative and generative phenotypes. With plants growing in the vegetative state, some LEACO1-ipt transgenic lines appeared similar to the non-transgenic wild-type cultivars while other lines showed excessive lateral branch development and short internodes. With plants grown under generative conditions, several LEACO1-ipt transgenic lines showed a 2 to 10-fold increase in the number of flower buds relative to the wild-type cultivars. With chrysanthemum, dramatic increases in bud count were observed on transgenic lines that otherwise displayed a morphology similar to the non-transgenic lines. Analysis of ipt expression indicated a marked change in gene expression between the most extreme phenotypes observed in this study. LEACO1-ipt lines that express normal vegetative development but increased flower bud counts appear to have great potential for ornamental crop improvement.

It has been reported that constitutive expression of the fatty acid desaturase enzyme increased the trienoic fatty acid content of thylakoid membranes in transgenic tobacco, allowing the membranes to remain fluid under cold conditions. While increased cold tolerance resulted from this genetic modification, plants with a constitutively expressed desaturase enzyme would not be particularly well suited for growth under warm temperatures. To increase the ability of plants to tolerate prolonged cold-storage and still perform under greenhouse production conditions (25 °C), a unique cold-inducible genetic construct was cloned and tested. The FAD7 gene, which encodes an omega-3-fatty acid desaturase enzyme, was put under the control of a cold-inducible promoter (cor15a) from Arabidopsis thaliana. Transgenic petunia plants (cv, Marco Polo Odyssey) harboring cor15a:FAD7 were established and conformed by PCR and Southern analysis. Therefore in our study, FAD7 gene expression was induced by exposure to cold temperatures and down regulated under normal growing conditions. RT-PCR indicated a marked increase in FAD7 expression between transgenic plants exposed to a short (3 days) cold treatment prior to long-term cold storage and those that did not receive a cold induction treatment. Transgenic and wild-type plants were induced in cold (3 °C) for 3 days, returned for normal greenhouse conditions for 5 days and then subjected 3 weeks of continuous cold storage. It was observed that two out of eight transgenic lines showed superior cold tolerance relative to wild-type petunia plants. Additionally, plants that showed cold tolerance completely recovered; growing and flowering normally when returned to the 25 °C greenhouse conditions.

The bacterial isopentenyl transferase (ipt) gene involved in cytokinin biosynthesis was fused with a promoter from the serine proteinase inhibiter (win3.12) gene of Populus x generosa and introduced into Nicotiana tabacum (cv. Havana). Transformants were confirmed by PCR reaction and Southern blot analysis, and then analyzed for phenotypic characteristics. RT-PCR analysis detected transcripts of the ipt gene following the wounding of win3.12:ipt transgenic plants. In win3.12: ipt transgenic plants, lateral shoot number and the diameter of lateral branches that developed following apical shoot removal increased relative to wild-type plants. However, the increase in cytokinin production following wounding appeared to be short lived. The potential utility of this construct in enhancing lateral branching in ornamental crops will be discussed.

The king oyster mushroom [Pleurotus eryngii (DC.:Fr.) Quél.] is gaining popularity across the world due to its excellent taste, high nutritional quality, medicinal value, and long shelf life. Conventional substrates for king oyster mushroom cultivation consist of sawdust derived from various tree species. Sawdust demand is increasing worldwide, creating a need for alternative materials that can at least partially replace sawdust as substrate for king oyster mushroom. In Taiwan, as in other countries that grow fruit trees, pruned fruit tree branches are an expensive agricultural waste, particularly if they are not recycled or reused. In the present study, we evaluated substrates containing sawdust and different proportions of material ground from pruned wax apple or Indian jujube branches for cultivation of king oyster mushroom. Our results suggested that among all five substrate mixes tested, the best substitute for conventional sawdust (100% sawdust) was a substrate that contained 75% sawdust mixed with 25% materials ground from trimmed wax apple branches (Wax apple 25%). Furthermore, determination of mineral element content, pH, and electrical conductivity (EC) levels of the substrates both before spawn inoculation and after harvesting revealed no significant changes in mineral content, a slight reduction in pH value, and a minor increase in EC levels after cultivation. Taken together, results from this study suggest that agricultural wastes from pruned fruit tree branches can partially replace sawdust as the cultivation substrate for king oyster mushroom.

Chill injury and leaf senescence occur in plants held in prolonged cold, dark storage. To increase tolerance to these conditions, Nicotiana alata and N. tabacum were transformed with either the FAD7 or IPT genes under the control of a cold-inducible promoter (cor15a). FAD7 encodes for omega-3-fatty acid desaturase and was used to resist cold-stress. IPT encodes the cytokinin-pathway enzyme isopentenyl transferase and was used to delay senescence. Independent FAD7 and IPT lines were crossed to produce double transgenic seed. Seedlings from single transgenic (cor15a-IPT or cor15a-FAD7) lines, double transgenic lines, and the wild-type were exposed to prolonged cold, dark conditions. After 3 months in the dark at 2 °C, survival of independent double transgenic N. tabacum lines ranged up to 80% to 90%. However only 40% of FAD7 seedlings survived, 10% of IPT seedlings survived, and no wild-type plants survived. Double transgenic N. alata seedlings average 90% survival under similar conditions and RT-PCR revealed expression of both the IPT and FAD7 genes. Omega-3-FAD enzyme activity increases desaturation in chloroplast membrane fatty acids. When exposed to prolonged cold, the molecular fraction of polyunsaturated fatty acids (18:3 and 16:3) in leaves of wild-type N. alata decreased while monounsaturated (16:1 and 18:1) and saturated fatty acid species (16:0 and 18:0) increased dramatically. In double transgenic N. alata lines exposed to prolonged cold, the molecular fraction of 18:3 and 16:3 increased, while the 16:0 and 18:0 species decreased dramatically compared to nonchilled double transgenic plants.

Cold stress is one of the most important environmental factors affecting crop growth and agricultural production. Induced changes of gene expression and metabolism are critical for plants responding and acclimating to cold stress. Banana (Musa sp.) is one of the most important food crops in the tropical and subtropical countries of the world. Banana, which originated from tropical regions, is sensitive to cold, which can result in serious losses in commercial banana production. To investigate the response of the banana to cold stress conditions, changes in protein expression were analyzed using a comparative proteomics approach. ‘Brazil’ banana (Musa acuminata AAA group) is a common banana cultivar in southern China. ‘Brazil’ banana plantlets were exposed to 5 °C for 24 hours and then total crude protein was extracted from treatment and control leaves by phenol extraction, separated with two-dimensional gel electrophoresis, and subsequently identified by mass spectrometry (MS). Out of the more than 400 protein spots reproducibly detected, only 41 protein spots exhibited a change in intensity by at least 2-fold, with 26 proteins increasing and 15 proteins decreasing expression. Of these, 28 differentially expressed proteins were identified by MS. The identified proteins, including well-known and novel cold-responsive proteins, are involved in several cellular processes, including antioxidation and antipathogen, photosynthesis, chaperones, protein synthesis, signal transduction, energy metabolism, and other cellular functions. Proteins related to antioxidation, pathogen resistance, molecular chaperones, and energy metabolism were up-regulated, and proteins related to ethylene synthesis, protein synthesis, and epigenetic modification were down-regulated in response to cold temperature treatment. The banana plantlets incubated at cold temperatures demonstrated major changes in increased reactive oxygen species (ROS) scavenging, defense against diseases, and energy supply. Increased antioxidation capability in banana was also discovered in plantain, which has greater cold tolerance than banana in response to cold stress conditions. Therefore, we hypothesized that an increased antioxidation ability could be a common characteristic of banana and plantain in response to cold stress conditions. These findings may provide a better understanding of the physiological processes of banana in response to cold stress conditions.

Understanding of root growth patterns and architecture of apple (Malus ×domestica Borkh.) trees is very important for commercial apple production. Most commercial apple trees are usually a grafted complex consisting of the scion and the rootstock, each of which is a different genotype. Recently, rootstocks of dwarf tree species have been used extensively to meet the convenience in management; however, this practice appears to negatively impact root development. Using minirhizotrons, we investigated root dynamics, root spatial distribution, and shoot growth in ‘Red Fuji’ scion grown: 1) directly on dwarf and vigorous root stocks and 2) on a dwarf root stock placed in between the non-dwarf scion and non-dwarf rootstock (hereinafter referred to as an interstem). The results showed that: 1) one or two peaks in total root length density (TRLD) were observed in each scion/rootstock combinations every year; 2) the greatest TRLD peaks were always observed in between May and December. The peaks of shoot growth were always asynchronous with that of white root length density; 3) compared with scion/vigorous rootstock combinations, inserting a dwarfing interstem between the scion and vigorous seedling rootstock reduced the TRLD; 4) scion/vigorous rootstock combinations had a relatively deep, widespread and large root system. Scion/dwarfing rootstock combinations had a root system distributed in a small region; and the root systems of scion/dwarfing interstem/vigorous rootstock combinations tended to be intermediate between those of scion/vigorous rootstock and scion/dwarfing rootstock. This implies that the insertion of interstems altered the root architecture by not only the quantity of roots, but also the spatial distribution.

Photosynthetic physiology of Dendrobium nobile, Dendrobium pendulum, Dendrobium chrysotoxum, and Dendrobium densiflorum was studied. A bimodal diurnal variation of the net photosynthetic rate (P_n) was observed in the four Dendrobium species with the first peak [5.09 to 6.06 μmol (CO₂) per m⁻²·s⁻¹] ≈1100 hr and the second peak [3.83 to 4.58 μmol (CO₂) per m⁻²·s⁻¹] at 1500 hr. No CO₂ fixation was observed at night. For all four Dendrobium species, the light compensation point (LCP) was 5 to 10 μmol·m⁻²·s⁻¹, light saturation point (LSP) ranged from 800 to 1000 μmol·m⁻²·s⁻¹, apparent quantum yield (AQY) was 0.02, and CO₂ compensation points (CCP) and saturation point (CSP) were 60 to 85 μmol·mol⁻¹ and 800 to 1000 μmol·mol⁻¹, respectively. Carboxylation efficiency (CE) values ranged from 0.011 to 0.020. The optimum temperature for photosynthesis was between 26 and 30 °C. The measurement of P_n seasonal variation indicated that July to August had the higher P_n for Dendrobium species. Additionally, the chlorophyll a/b (Chl a/b) ratios of the leaves were 2.77 to 2.89. Measurement of key enzymes in the photosynthetic pathway indicated relatively high Ribulose-1,5-bisphosphate carboxylase (RuBPCase) and glycolate oxidase (GO) activities but very low phosphoenolpyruvate carboxylase (PEPCase) activities. It suggested that these four Dendrobium species are typical semishade C₃ plants.