Overexpression of TaLEA3 induces rapid stomatal closure under drought stress in Phellodendron amurense Rupr.

Late embryogenesis abundant (LEA) proteins participate in drought stress responses in plants. In the present study, the gene TaLEA3 from the drought-resistant plant Tamarix androssowii was transformed into Amur cork tree (Phellodendron amurense) via Agrobacterium tumefaciens to investigate the mechanism of stomatal closure in response to osmotic stress. Our results showed that P. amurense overexpressing TaLEA3 were resistant to drought stress by rapid stomatal closure. To study the stomatal movement regulated at the molecular level, a model system for stoma closure was established in in vitro P. amurense. In this work, we found that the increased Ca2+ accumulation in guard cells of transgenic plants caused stomatal closure and activated K+ efflux under polyethylene glycol (PEG) stress. Moreover, H+ changes might provide a needed pH condition for stomatal closure. Further, nitric oxide (NO) fluorescence was measured using an NO-specific fluorescent probe, diaminofluorescein-FM diacetate, which showed that guard cell NO fluorescence was stronger in transgenic plants compared with wild type plants. Additionally, five genes encoding nitrate reductase were up-regulated, indicating that TaLEA3 overexpression positively regulated NO biosynthesis and accumulation in the guard cells. This discovery will further our understanding of the LEA gene function and will help in engineering drought-resistant tree cultivars.

[Effects of light quality on the seed germination of main tree species in a secondary forest ecosystem of Northeast China].

This paper explored the effects of light quality on the seed germination of five dominant tree species (Larix kaempferi, Phellodendron amurense, Acer mono, Fraxinus mandshurica, and Pinus koraiensis) in a secondary forest ecosystem of Northeast China, based on the experiments with the seeds of the five tree species in laboratory and those of the P. koraiensis and L. kaempferi in the field. Four treatments of different light quality were designed in laboratory (taking dark as the control), and three treatments of R/FR (the ratio of red light and far red light intensity) were installed in the field. The laboratory experiment showed that light quality had less effect on the seed germination of L. kaempferi, but the seed germination rates of the other four tree species were significantly different under the treatments of different light quality. P. amurense had the highest seed germination rate under white light, whereas A. mono, F. mandshurica, and P. koraiensis had the highest one under the alternative irradiation with red light and far red light (R-FR-R). In consistence with the results in laboratory, the seed germination rate of P. koraiensis in the field decreased with decreasing R/FR ratio, while that of L. kaempferi was less affected. Under natural condition, the R-FR-R fluctuated with the activity of sun-fleck, and the seed germination patterns of A. mono, F. mandshurica, and P. koraiensis could be the adaptation to the sun-fleck environment in forest stand. The germination of large seeds was significantly affected by light quality.

[Ecological evaluation of suitable area for production of Phellodendron amurense based on TCMGIS- I].

The TCMGIS- I (Suitability evaluation geographic information system of traditional Chinese medicine producing area ) was developed and used to analyze the appropriate producing area of Phellodendron amurense in China. Taoshan county of Heilongjiang province, one of the main producing areas of the geo-authentic crude drug, was selected as the analytical basal place. The results show that besides traditional producing area in Taoshan county of Heilongjiang province, 131 counties of the provinces of Heilongjiang, Jilin, Liaoning and Innermongolia are also alternative areas according to the eco-soil condition that the medicinal plants needed, and the sum area are 567 989. 3 m2, the results also indicate some condign regions in Innermongolia municipality. The system is much valuable to the division of adaptive area, introduction and acclimatization of medicinal materials.

[Construction and analysis of subtractive cDNA library of Phellodendron amurense under drought stress].

With cDNA from Phellodendron amurense seedlings treated with drought stress as tester and cDNA from this plant in normal growth as driver, we construct cDNA subtracted library using suppression subtractive hybridization (SSH). In the library, the rate of recombination was 95%, the size of inserts was 300-800 bp. Two hundred and sixty-five new genes were obtained by DNA sequencing 816 positive clones picked randomly, and partitioned to 16 classes after nucleotide Blast and BlastX homological analysis against NT, NR, SWISSPROT, KEGG database. Forty-four drought stress associated genes, such as heat shock protein cognate 70, dehydration responsive protein 22, universal stress protein, metallothionein II, late embryogenesis abundant protein, were obtained, which made 16.6% of the overall genes. These genes included osmotic regulator, signal component regulatory protein and antioxidant enzyme. The research had established a basis for cloning stress resistance genes and further studying genes expression in P. amurense seedlings under drought stress.