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  • Author or Editor: Long Chen x
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Identification of low temperature–regulated gene expression in Pachysandra terminalis: Pachysandra terminalis is a cold-hardy, evergreen plant species. In order to identify molecular mechanism of cold tolerance of this plant species, seedlings with four fully expanded leaves were subjected to 4, 0, and –1 °C low temperature treatments. Low temperature–induced genes were identified from treated plants using cDNA differential display. The cDNA fragments were cloned onto PCR-trap vectors. Low temperature regulation of these genes was confirmed by reverse-northern blot. Sequence analysis has identified that these genes can be classified into three groups, stress-related, photosystem-related. Most of the genes cannot find matching sequences in the database. To further study the regulation of these genes by temperature fluctuation, the plants were treated at 4, 0, and 40 °C. Northern blot analysis showed that several clones showed increased expression after cold and heat shock. Previous cold treatment at 4 °C can negate the effect of heat shock on expression of these genes. Complete sequence of these genes is cloned from the cDNA library and their temporal regulation by environmental stresses is analyzed using real-time PCR.

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Chimonanthus praecox (wintersweet) is endemic to China. It has been cultivated there for more than 1000 years as a garden, potted, and cut-flower plant. Many cultivars have been developed during its long history of cultivation, and recently many germplasms were collected in Wuhan and Nanjing, China. The identification and genetic relationship of these resources were studied based mainly on morphological traits. In the current study, intersimple sequence repeat markers (ISSR) and random amplified polymorphic DNA markers (RAPD) were used for the first time to investigate 72 wintersweet clones from the two regions. Eleven ISSR primers amplified 115 bands, 90 (78.26%) of which were polymorphic. Nineteen RAPD primers amplified 165 bands, 105 (63.63%) of which were polymorphic. Either ISSR or RAPD markers were sufficient to distinguish all the clones surveyed. A Dendrogram based on Jaccard's similarity coefficients indicated that the distribution pattern of the 72 clones was coherent with their geographical origins. Most of the genetic variation (85.68% with ISSR data; 86.75% with RAPD data) occurred among clones within each region. However, the difference between Wuhan and Nanjing groups is statistically significant (ΦST = 0.143, P < 0.001, with ISSR data; ΦST = 0.132, P < 0.001, with RAPD data). Morphological variation and classification of wintersweet cultivars were also discussed compared with the genetic relationship based on ISSR and RAPD markers. This is the first report of the partitioning of genetic variability within and between different cultivated wintersweet regions, and it provides useful baseline data for optimizing sampling strategies in breeding. These results are important for future genetic improvement, identification, and conservation of Chimonanthus praecox germplasm.

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Shennongjia mountain region is famous for its various kinds of species. Through one year's deep-going expedition in the area, lots of valuable plant species were collected, among them many are very useful and had not been used in landscape. Such as Arisaema lobatum var. variegatum nv. LuDiFei, Cremastra appendiculata var. fulva LuDiFei, Stylophorum lasiocarpum (Oliv.) Fedde, Sedum filipes Hems., Iris wilsonii C. H. Wright, Amaranthus caudatus L., Cotoneaster dammeri Schneid, Meconopsis quintupineria Regel., Lysimachia paridiformis Franch., Dysoma versipellis (Hance) M. Cheng, Adiantum pedatum L. and so on. Some genera are quite rich in this region, especially in Rosa, Sorbus, Cotoneaster, Lonicera, Impatiens, Aconitum, Gentiana, Adiantum etc. All these are marvelous material for direct appliance in garden and for breeding. There are many rare plants in the area, large communities of Davidia involucrata Baillon and Chimonanthus praecox (L.) Link were found during the expedition, and what interesting more is that various natural variations do exist in the communities. Detail description and evaluation were given to the important species, and some suggestions of protection and utilization were offered in the paper.

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Ten polymorphic microsatellite loci were isolated and characterized from an enriched genomic library of Paphiopedilum concolor (Batem.) Pfitzer. The number of alleles per microsatellite locus ranged from three to 11 with an average of 6.4 in a sample of 30 individuals from three populations. The observed and expected heterozygosity ranged from 0.200 to 0.800 and from 0.544 to 0.827, respectively. These microsatellites can be used as tools to investigate the genetic structure of P. concolor populations and relationship patterns with closely related taxa.

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Dendrobium officinale Kimura et Migo is a famous traditional Chinese medicinal plant. It produces various phytochemicals, particularly polysaccharides, which have nutraceutical and pharmaceutical values. To increase its biomass production and polysaccharide content, our breeding program has generated a series of polyploid cultivars through colchicine treatment of protocorm-like bodies (PLBs). The present study compared two tetraploid cultivars, 201-1-T1 and 201-1-T2, with their diploid parental cultivar, 201-1, in an established in vitro culture system. Tetraploid ‘201-1-T1’ and ‘201-1-T2’ had shorter leaves and shorter and thicker stems and roots, and they produced higher biomass compared with the diploid cultivar. The length and width of stomata significantly increased, but stomatal density decreased in tetraploid cultivars. The PLB induction rates from the stem node explants of the tetraploid cultivars were significantly higher than those of diploid. However, the PLB proliferation of tetraploids was lower than that of the diploid. The mean number of plantlets regenerated from tetraploid PLBs was also lower than that of the diploid after 4 months of culture. Polysaccharide contents in stems, leaves, and roots of 6-month-old tetraploid plantlets were significantly higher than those of diploids. The polysaccharide content in the stem of ‘201-1-T1’ was 12.70%, which was a 2-fold increase compared with the diploid cultivar. Our results showed that chromosome doubling could be a viable way of improving D. officinale in biomass and polysaccharide production.

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