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In Pelargonium, the plastid mutation in three independent cell layers L1, L2, and L3, can produce plastid chimeras with visible shoot colour difference such as GWG (green-white-green) and GGW (green-green-white). Chimera can be used to trace the relationship between the cell layers of different genotypes during shoot development and the effect of the mutated genes on shoot development. In this study, we have obtained different adventitious shoots with GGG, GWG, GGW, and WWW combinations of cell layers through tissue culture of petioles and internodes from GGW and GWG chimeras of Pelargonium zonale `Mrs Pollock'. Much higher percentage (14.9%) of chimeral adventitious shoots was obtained from GGW tissues than from GWG tissues (4.2%). Of the 10.8% chimeral adventitious shoots regenerated in this experiment, 8.6% are different from the original type of explants. This result indicated that cells at least in both L2 and L3 of the explants were involved in the regeneration of the adventitious shoots. The number of shoot types regenerated is likely dependent on the number and the type of cells that were in direct contact with the culture medium. It is suggested that the mixed cells can be used to produce the chimera by tissue culture. Three possible ways to form the chimeras in vitro culture were discussed. Chemical names used: TDZ =1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea (Thidiazuron); IAA = Indole-3-acetic acid; PVP = polyvinylpyrrolidone.

Sixty highly homozygous tomato (Lycopersicon esculentum Mill.) strains, some selected from previous studies and some collected from known low-Ca regions, were screened under a low-Ca culture system (10 mg of Ca per plant). Four strains were selected to represent the extremes for Ca efficiency and used as parents to create a series of F₁, F₂, and backcross generations for inheritance studies of Ca use under low-Ca stress. Based on total plant dry weight, additive and dominance gene effects were most important for the efficiency of Ca use. Maternal control of efficiency in Ca use was not observed. Estimates of broad-sense heritability ranged from 63% to 79% for total play dry weight. Narrow sense heritabilities, determined in only two of the families, were 47% to 49$ and 68% to 75%.

Long-term exposure to an elevated ambient carbon dioxide (eCO₂) concentration could weaken or diminish the enhancement of plant photosynthesis and growth. To monitor this response and offer references for growth management, the whole-plant photosynthetic rate (P _n,w ) and dark respiration rate (R _d,w ) of Angelica acutiloba Kitagawa transplants were monitored with a growth chamber. The results showed that eCO₂ increased both the P _n,w and R _d,w by (79 ± 42) % and (126 ± 51) %. The dry weight of transplants under eCO₂ was 33.6% greater than that under aCO₂. However, the photosynthetic acclimation to eCO₂ occurred. The increase in the P _n,w was maintained until the end of the experiment due to increased leaf area. Moreover, the increase in plant dry weight mainly occurred in the first 15 days of treatment. Furthermore, the dry weight estimated based on the P _n,w and R _d,w agreed well with the measured dry weight. The relative growth rate (RGR) calculated with the estimated dry weight demonstrated the response of transplant growth to eCO₂. These results indicated that the proposed method can be used to monitor the response of plant growth to eCO₂.

The Himalayan yew, Taxus wallichiana Zucc., is an endangered species with a scatted distribution in the Eastern Himalayas and southwestern China. In the present study, 10 microsatellite markers from the genome of T. wallichiana were developed using the protocol of fast isolation by amplified fragment length polymorphism of sequences containing repeats (FIASCO). Polymorphism of each locus was assessed in 28 samples from four wild populations of the Himalayan yew. The allele number of the microsatellites ranged from two to five with an average of 2.9 per allele. The observed and expected heterozygosity varied from 0.00 to 1.00 and from 0.3818 to 0.7552, respectively. Cross-species amplification in another two yew species showed eight of them holding promise for sister species. Two of the 10 loci (TG126 and TC49) significantly deviated from Hardy-Weinberg expectations. No significant linkage disequilibrium was detected between the comparisons of these loci. These polymorphic microsatellite markers would be useful tools for population genetics studies and assessing genetic variations to establish conservation strategy of this endangered species.

Apple blotch caused by Alternaria alternata apple pathotype is a severe disease of apple (Malus ×domestica Borkh) occurring throughout the world, especially in eastern Asia. Phenotypic and genetic information about resistance/susceptibility of apple germplasm to this disease will be extremely valuable for selecting and developing new disease resistant cultivars. In this study, 110 apple cultivars obtained from the USDA apple germplasm in Geneva, NY, were evaluated for their resistance/susceptibility to apple blotch by field surveys, and inoculation of detached leaves with a suspension of germinated conidia of A. alternata apple pathotype. Disease incidence were different among the cultivars and categorized into resistant (R), moderately resistant (MR), or susceptible (S). Two molecular markers, S428, a random amplified polymorphic DNA (RAPD) marker associated with disease resistance, and a simple sequence repeat (SSR or microsatellite) marker CH05g07, linked to susceptibility were used to correlate the phenotypes expressed in field surveys and laboratory inoculations. The detection using either the S428 marker or the CH05g07 marker in 50 common breeding cultivars was consistent with R or S traits except for ‘Bisbee’ and ‘Priscilla’. These two cultivars were MR to apple blotch through phenotyping. However, SSR markers were detected, but RAPD markers were not and therefore were considered susceptible. Combined with the record of resistance to fire blight from Germplasm Resources Information Network (GRIN), ‘Dayton’, ‘Mildew Immune Seedling’, ‘Puregold’, and ‘Pumpkin Sweet’ were highly resistant to both diseases and considered as the best choices of parents for stacking resistance to multiple diseases in breeding program.

Alternaria alternata apple pathotype (previously A. mali) causes alternaria blotch disease of apple (Malus ×domestica), which may result in leaf spots and up to 70% premature leaf drop in serious cases. This disease is of worldwide importance but is most serious in eastern Asia (Japan, Korea, and China) and in parts of the United States. The excessive use of fungicides not only adds cost to apple growers, but also pollutes the environment. In this study, we characterized a 5-year F₁ population from a cross of a resistant cultivar (Huacui) and a susceptible cultivar (Golden Delicious) consisting of 110 individuals along with 14-year-old parent trees (10 each). A field evaluation of disease severity was conducted in 2008 and 2009 under the natural conditions in Liaoning, China (lat. 40°37′ N, long. 120°44′ E). Based on the field data, 110 F₁ plants were divided into five groups. Artificial inoculation was carried out both on the living trees and on the detached leaves in 2009 to ensure that A. alternata apple pathotype was the causative agent. Eighty primer pairs of simple sequence repeat (SSR) were screened against the four genomic DNA pools, respectively, from six highly susceptible F₁ plants, six most resistant F₁ plants, one tree of the seed parent, and the one tree of the pollen parent. One pair of primers (CH05g07) was shown to be linked to the DNA pools of susceptible F₁ and the parent tree, but not to the DNA pools of resistant F₁ and parent trees. This primer pair was then used to screen all individual 110 F₁ progenies and two parent trees. The differentiation of 103 individuals (97.3%) with the marker matched the field disease resistance rating. This marker was further screened with 20 cultivars with known susceptibility or resistance to A. alternata apple pathotype and its linkage to susceptibility was validated. These results suggest that this marker can be used in marker-assisted selection for resistance/susceptibility to alternaria blotch disease in apple.

Pyracantha coccinea is a thorny evergreen shrub native to southeast Europe to southeast Asia. It is a popular ornamental plant because of its showy bright red fruits and small white flowers. However, in vitro vegetative propagation of P. coccinea has not been studied. Nodal segments with one or two axillary buds (1 to 1.5 cm in length) were cut and disinfected in a solution of 0.1% (v/v) mercuric chloride (HgCl₂) for 5 minutes, and proliferated on Murashige and Skoog (MS) basal medium supplemented with various concentrations 6-benzylaminopurine (6-BA). After 4 weeks, newly formed shoots were transferred to proliferation and rooting media containing various concentrations of indole-3-butyric acid (IBA). Establishment of axillary buds was significantly better with an establishing rate of 67% on basal MS medium augmented with 6.6 µm 6-BA. The best medium for proliferation of shoots was three-fourth basal MS supplemented with 1.5 µm IBA, with a proliferation rate of 3.4 axillary bud. The optimum rooting medium was one-fourth MS basal medium containing 93 µm IBA. Rooting of shoots was as much as 77%. Rooted plantlets were transferred to pots containing vermiculite:perlite:peat (6:1:2) and acclimatized to ambient greenhouse conditions with a 95% survival rate. This protocol can be used for in vitro propagation of P. coccinea.

Nitrogen and potassium are two crucial nutrient elements that affect the yield and quality of crops. The aim of this study was to quantify the impacts of potassium on growth dynamics and quality of muskmelon, so as to optimize potassium management for muskmelon in a plastic greenhouse, and develop a coupling model of nitrogen and potassium. For this purpose, four experiments (two experiments with different levels of potassium treatment and planting dates, and the other two experiments with different ratios of nitrogen and potassium, and planting dates) on muskmelon (Cucumis melo L. ‘Nanhaimi’ and ‘Xizhoumi 25’) were conducted in a plastic greenhouse located at Sanya from Jan. 2014 to Sept. 2015. The quantitative relationship between leaf potassium content and growth dynamics and yield of muskmelon was determined and incorporated into a photosynthesis-driven crop growth model (SUCROS). Independent experimental data were used to validate the model. The critical leaf potassium content at the flowering stage for muskmelon ‘Nanhaimi’ and ‘Xizhoumi 25’ were 55.0 and 46.0 mg·g⁻¹. The result showed that the coefficient of determination (r ²) between the predicted and measured values of leaf area index (LAI), direct weight of shoot (DWSH), direct weight of stem (DWST), dry weight of leaf (DWL), dry weight of fruit (DWF), fresh weight of fruit (FWF), soluble sugar content (SU), soluble protein content (PR), vitamin C (Vc), and soluble solids content (SO) of potassium model were 0.93, 0.98, 0.83, 0.96, 0.98, 0.99, 0.94, 0.94, 0.89, 0.85, and 0.90, respectively; and the relative root-mean-squared error (rRMSE) were 10.8%, 19.6%, 30.3%, 21.1%, 11.9%, 17.2%, 13.9%, 27.8%, 20.6%, and 10.1%, respectively. The two ways of nitrogen and potassium coupling (multiplicative coupling and minimum coupling) were compared, and the multiplicative coupling was used in model development finally. The r ² between the predicted and measured values of LAI, DWSH, DWST, DWL, DWF, FWF, SU, PR, Vc, and SO of nitrogen and potassium coupling model were 0.78, 0.91, 0.93, 0.94, 0.83, 0.89, 0.92, 0.95, 0.91, and 0.93, respectively; and their rRMSE were 9.2%, 12.4%, 11.8%, 43.2%, 6.6%, 7.2%, 6.85%, 4.98%, 6.61%, and 4.35%, respectively. The models could be used for the optimization of potassium, nitrogen, and potassium coupling management for muskmelon production in a plastic greenhouse.

Seasonal alteration of the cytosolic and nuclear Ca²⁺ concentrations of spruce (Picea engelmannii Parry) and brome grass (Bromus inermis Leyss) was investigated by the antimonate precipitation cytochemical technique. Electron microscopic (EM) observations revealed that electron-dense Ca²⁺ antimonate deposits, an indication of Ca²⁺ localization, were seen mainly in the vacuole, the cell wall and the intercellular space in samples of both species, collected on 14 July 1997. Few deposits were found in the cytosol and nuclei, showing a low resting level during summer months. On 8 Aug. 1997 following a decrease in daylength of 1 hour and 12 minutes, Ca²⁺ accumulation was initiated in spruce with increased cytosolic and nuclear Ca²⁺ deposits, but not in brome grass. On 8 Sept. 1997, Ca²⁺ accumulation occurred in the cytosol of brome grass. This followed a drop in ambient temperature to 12 °C. Cytosolic and nuclear Ca²⁺ deposits continued to increase in spruce. Controlled experiments confirmed that it was the low temperature, not shortening daylength, that triggered Ca²⁺ accumulation in brome grass. High cytosolic and nuclear Ca²⁺ concentrations lasted about three months in spruce from early August to early November. However, the high cytosolic and nuclear Ca²⁺ concentrations in brome grass lasted only about 20 days from early September to the end of the month. During winter and spring, both species had low resting cytosolic and nuclear Ca²⁺ concentrations. The relationship between the duration of the high cytosolic and nuclear Ca²⁺ concentrations and the status of the developed dormancy/cold hardiness is discussed in light of current findings.

Chinese cymbidiums (Cymbidium sp.) are important ornamental plants because of their foliage, flower shape, and fragrance. Well-known Chinese cymbidiums mainly include Cymbidium goeringii, Cymbidium faberi, Cymbidium ensifolium, Cymbidium kanran, and Cymbidium sinense. The population genetics of Chinese cymbidiums can be efficiently analyzed using small-scale marker panels with high discriminatory power. In this study, we tested several genic simple sequence repeats (SSRs) and built six genic SSR panels. The panels included several robust markers, which can rapidly assign Chinese cymbidium accessions to their source species. Fifty-three accessions of Chinese cymbidiums were analyzed using 25 markers, which exhibited polymorphism among five species. These markers were ranked according to their discriminatory scores (D scores). The program selected six markers to build an “overall” panel for all Cymbidium classifications and yielded 95.16% population assignment accuracy. Considering one species as the “critical” population and the four other species as one population, we built five genic SSR panels: C. ensifolium panel (four markers, 98.05% accuracy), C. faberi panel (six markers, 95.90% accuracy), C. goeringii panel (six markers, 95.15% accuracy), C. sinense panel (six markers, 96.35% accuracy), and C. kanran panel (five markers, 96.10% accuracy). Genetic distance matrices calculated using the “overall” panels and those derived with the 25 markers were compared. Results showed a high correlation (R = 0.807) with statistical significance (P = 0.042). Moreover, “all panels” revealed higher genetic variations among populations than “all markers.” Hence, the developed panels are suitable for efficient population classification of Chinese cymbidiums.