Characterization and Expression Analysis of Four Glycine-Rich RNA-Binding Proteins Involved in Osmotic Response in Tobacco (Nicotiana tabacum cv. Xanthi)

Plants have developed many signals and specific genes＇ regulations at both transcriptional and post-transcriptional levels in order to tolerate and adapt to various environmental stresses. RNA-binding proteins （RBPs） play crucial roles in the post- transcriptional regulation via mRNA splicing, polyadenylation, sequence editing, transport, mRNA stability, mRNA localization, and translation. In this paper, four cDNAs of glycine-rich RNA-binding proteins （GR-RBPs）, named NtRGP-la, -lb, -2, and -3, were isolated from Nicotiana tabacum by RT-PCR analysis, and special emphases were given to the sequences alignment, phylogenetic analysis and gene expression. Sequences alignment revealed minor difference of cDNA sequences, but no difference of deduced proteins between N. sylvestris and N. tabacum. Phylogenetic alignment revealed that four cDNAs in tobacco were clustered into two different groups. NtRGP-2 and -3 were evolutionarily closest to Arabidopsis GR-RBPs genes and related to animal GR-RBPs genes, while NtRGP-la and -lb were closest to Gramineae GR-RBPs genes. The expression analyses of these four NtRGPs in response to different abiotic stresses revealed the similar expression pattern. Moreover, the four NtRGPs, especially NtRGP-la and NtRGP-3, were strongly induced by stresses including water, wound, cold, and high temperature, weakly induced by PEG, drought and SA, while reduced by NaC1 and unaffected by ABA treatment. The fact that all of these abiotic stresses included in our experiments affected the water balance and resulted in osmotic stress on cellular level, suggests that NtRGPs in tobacco should be a family of crucial osmosis-related proteins, and may play a key role in signal transduction with ABA-independent pathway under abiotic stresses.

Effects of Different Flue-curing Technologies on Submicroscopic Structure of Flue-cured Tobacco Leaves

[ Objective] This study aimed to compare the effects of low-temperature low-humidity tobacco flue-curing technology, moderate-temperature moderate- humidity tobacco flue-curing technology and moderate-temperature high-humidity tobacco flue-curing technology on submicroscopic structure of flue-cured tobacco leaves to provide theoretical basis for tobacco flue-curing. [ Method] Middle leaves of tobacco cuhivar K326 were collected in Pengshui County of Chongqing City for flue-curing experiment using three flue-curing technologies. Leaf samples were collected regularly in the flue-curing process, to investigate the submicroscopic structure of flue-cured tobacco leaves. [ Result] Morphology changes and structural rupture of chloroplasts occurred at different time during three flue-curing proces- ses. In low-temperature low-humidity flue-curing process, morphology changes and structural rupture of chloroplasts occurred late but rapidly; in moderate-tempera- ture moderate-humidity flue-curing process and moderate-temperature high-humidity flue-curing process, morphology changes and structural rupture of chloroplasts occurred early but slowly. Among these three flue-curing processes, the speed of changes in mitochondria morphology, cristae number and membrane rupture pres- ented an upward order of low-temperature low-humidity flue-curing process 〈 moderate-temperature high-humidity flue-curlng process 〈 moderate-temperature moderate-humidity flue-curing process. In low-temperature low-humldity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 36 -48 h ; in moderate-temperature moderate-humidity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 24 - 36 h; in moderate-temperature high-humidity flue-curing process, starch granules in chloroplasts ruptured after flue-curing for 12 - 24 h. [ Conclusion ] Using low-temperature low-humidity flue- curing technology could prolong the maintenance duration of the submicroscopic structure integrity of flue-cured tobacco leaves, which was conducive to fully decom- posing and transforming substances in tobacco leaves and obtaining high-quality flue-cured tobacco leaves.

Effects of Different Flue-curing Technologies on Microscopic Structure of Flue-cured Tobacco Leaves

[Objective] This study aimed to compare the effects of Chinese quality-improving fragrunce-increasing tobacco flue-curing technology, Chinese three- stage tobaeeo flue-curing technology and Zimbabwean tobaceo flue-euring technology on microscopie structure of flue-cured tobacco leaves to provide theoretical basis for tobacco flue-curing. [ Method ] Middle leaves of tobacco cultivar K326 were collected in Pengshui County of Chongqing City for flue-curing experiment using three flue-curing technologies. Leaf samples were collected regularly in the flue-curing process, to investigate the microscopic structure of flue-cured tobaceo leaves. [ Result] During three flue-curing processes, leaves, palisade tissues and sponge tissues shrank gradually. Three flue-euring processes exhibited significant differ- ences in the peak of tissue shrinkage ： microscopic structure of flue-cured tobacco leaves in Chinese three-stage tobacco flue-curing process shrank earliest, follower by Chinese quality-improving fragrance-increasing tobacco flue-curing process; flue-eured tobacco leaves in Zimbabwean tobacco flue-curing process presented the latest shrinkage. At 84 h post-curing, leaf thickness in three flue-curing processes showed a descend order of Chinese three-stage tobacco flue-curing process 〉 Chinese quality-improving fragrance-increasing tobacco flue-curing process 〉 Zimbabwean tobacco flue-curing process. Upper and lower epidermal cells in Zimb- abwean tobacco flue-curing process ruptured earlier than other two flue-curing processes; eventually, the majority of cells ruptured and mixed with palisade tissues and sponge tissues. In Chinese quality-improving fragranee-increasing tobacco flue-curing process, only a small number of epidermal cells in dried leaves ruptured. Among three flue-curing technologies, Zimbabwean tobacco flue-curing technology exhibited the greatest damage to epidermal cells, followed by Chinese three-stage tobaceo flue-curing technology; Chinese quality-improving fragrance-increasing tobacco flue-curing technology had the minimum damage to epidermal cells. Mese- phyll cross-section exhibited significant morphological changes in stomata. To be specific, at 0 - 12 h post-curing, stomata of tobacco leaves closed with slightly prominent stamatal apparatuses on upper and lower epidermis; at 24 -72 h post-curing, stomata of tobaceo leaves changed gradually from opening to closure with significantly prominent stomatal apparatuses on upper and lower epidermis; at 84 h post-curing, stomata of tobacco leaves closed, and the majority of stomatal appa- ratuses were significantly prominent. In Chinese quality-improving fragrance-increasing tobacco flue-curing process, only a small number of epidermal ceils ruptured at 84 h pest-flue-curing; palisade tissues and sponge tissues shrank almost simultaneously. Significant gaps were observed between palisade tissues and between pal- isade tissues and sponge tissues. Chinese quality-improving fragrance-increasing tobacco flue-curing technology exhibited lower disorder level compared with other two flue-curing technologies. [ Conclusion] Chinese quality-improving fragrance-increasing tobacco flue-curlng technology was conducive to maintaining the micro- scopic structure integrity of flue-cured tobacco leaves and obtaining high-quality flue-cured tobacco leaves.

QPSO-optimized BP Neural Net ork to Predict Occurrence Quantity of Myzus persicae

In order to effectively predict occurrence quantity of Myzus persicae, BP neural network theory and method was used to establish prediction model for oc- currence quantity of M. persicae. Meanwhile, QPSO algorithm was used to optimize connection weight and threshold value of BP neural network, so as to determine. the optimal connection weight and threshold value. The historical data of M. persica quantity in Hongta County, Yuxi City of Yunnan Province from 2003 to 2006 was adopted as training samples, and the occurrence quantities of M. persicae from 2007 to 2009 were predicted. The prediction accuracy was 99.35%, the mini- mum completion time was 30 s, the average completion time was 34.5 s, and the running times were 19. The prediction effect of the model was obviously superior to other prediction models. The experiment showed that this model was more effective and feasible, with faster convergence rate and stronger stability, and could solve the similar problems in prediction and clustering. The study provides a theoretical basis for comprehensive prevention and control against M. persicae.

Analysis of Differential Gene Expression Pattern in Brassica napus Hybrid Huayouza 6 and Its Parents Using Arabidopsis cDNA Microarray

Huayouza 6, a new semi-winter Brassica napus variety with high-yield, good quality, prematufity and extensive adaptability, was derived from the cross between the female parent 8086A and male parent 7-5. Two cDNA-based Arabidopisis microarray were used to analyze gene differential expression in bud of an elite B.napus hybrid Huayouza6 and its parents, in which there were 83 over-expression transcripts and 331 under-expression transcripts between Huayouza 6 and its female parent 8086A and 94 over-expression transcripts, and 423 under-expression transcripts were demonstrated between Huayouza 6 and its male parent 7-5. Further analysis showed that there were significant number of genes responsible for photosynthesis, and its implication for heterosis was discussed. Northern analysis of phosphoribulokinase coincided with its expression pattern derived from hybridization of Arabidopsis cDNA microarray and B. napus mRNA, this system of heterologous hybridization analysis should be applicable to other close relatives of Arabidopsis thaliana.

Effects of Elevated CO_2 on Growth, Carbon Assimilation, Photosynthate Accumulation and Related Enzymes in Rice Leaves during Sink-Source Transition

To study the effects of growing rice （Oryza sativa L.） leaves under the treatment of the short-term elevated CO2 during the period of sink-source transition, several physiological processes such as dynamic changes in photosynthesis, photosynthate accumulation, enzyme activities （sucrose phosphate synthase （SPS）, and sucrose synthase （SS））, and their specific gene （spsl and RSusl） expressions in both mature and developing leaf were measured. Rice seedlings with fully expanded sixth leaf （marked as the source leaf, L6） were kept in elevated （700 μmol/mol） and ambient （350 mol/L） CO2 until the 7th leaf （marked as the sink leaf, L7） fully expanded. The results demonstrated that elevated CO2 significantly increased the rate of leaf elongation and biomass accumulation of L7 during the treatment without affecting the growth of L6. However, in both developing and mature leaves, net photosynthetic assimilation rate （A）, all kinds of photosynthate contents such as starch, sucrose and hexose, activities of SPS and SS and transcript levels of spsl and RSusl were significantly increased under elevated CO2 condition. Results suggested that the elevated CO2 had facilitated photosynthate assimilation, and increased photosynthate supplies from the source leaf to the sink leaf, which accelerated the growth and sink-source transition in new developing sink leaves. The mechanisms of SPS regulation by the elevated CO2 was also discussed.