Abstract:Seeking monoclone in established cDNA library randomly for PCR directed with universal M13 primer and sequencing,comparing the sequence in GenBank,designing specific primer according to the sequence,finally we obtained the whole ORF of siCOR.Then plant expression vector pBIN438-siCOR was constructed and NC89 tobacco plants were transformed via Agrobacterium tumefaciens,RT-PCR analysis indicated that siCOR had been integrated into the genome.Homozygous transgenic tobacco plants were used in cold resistance identification.The results show that:(1)The size of siCOR cDNA was 661 bp,the ideutity of amino acid sequence was about 65.04% with Oryza sativa Japonica.(2)siCOR tobacco has deep green leaves,grows faster and has no lateral bud compared with wild type;Under the cold stress,the decrease of relative water content,reduction of relative quantum yield of PSⅡ,the increase of relative conductivity and MDA content of siCOR tobacco are lesser than that of control tobacco evidently.Besides,transgenic plants have slower and lighter symptom.In summary,under the cold stress,siCOR tobacco shows well growth traits and strong cold resistance characteristics.

Abstract:Polygalacturonase-inhibiting protein (PGIP) can inhibit the activity of pathogen PG and improve the plant resistance level.(1)A novel PGIP gene designated as CsPGIP was isolated and characterized in Cucumis sativus L.The cDNA of this gene was 1 026 bp with an open reading frame of 987 bp encoding a 328-animo acid polypeptide.The deduced amino acid sequence of the gene contained the typical domain of xxLxLxxNxLt/sGxIPxxLxxLxxL and belonged to Pgip gene family.(2)Phylogenetic analysis revealed the significant evolutionary homologous in the sequences of melon PGIP gene and PGIP genes in Cruciferae.(3)Real-time quantitative PCR analysis showed that CsPGIP transcripts were expressed in all tissues,but the transcripts were most abundantly expressed in the young leaf but much less in stem.The expression level of the mRNA could be up-regulated significantly after treated with pathogen and salicylic acid (SA) compared to the normal growth environment.The results indicated that CsPGIP might play an important role on host resistance against pathogen.

Abstract:In this study,a promoter fragment of the SCBP60g gene was amplified by PCR from Arabidopsis,and fused with the GUS reporter gene.The recombination construct was then transformed into wild type Arabidopsis.Histochemical staining of GUS activity showed that:the promoter could drive the GUS expression in root,stem,leaf and flower,and relatively high expression levels were found in vascular tissue.However,this expression pattern is different from the promoter of LCBP60g that drives the same GUS gene expression,indicating the specific regulation of this promoter.

Abstract:A full-length cDNA of COMT,which is closely related to the biosynthesis of galanthamine in Amaryllidacea plants,was first successfully cloned from Lycoris radiata by the method of homology cloning and RACE,and was named as LrCOMT.The nucleotide analysis showed that the cDNA sequence of LrCOMT gene had 1 246 bp,containing a 1 101 bp open reading frame,which encodes 366 amino acid residues.Amino sequence analysis demonstrated that the deduced amino sequence of LrCOMT has many typical features of COMTs,and shared more than 60% indentity with COMTs from Iris hollandica,Zea mays,Nicotiana tabacum and other plants.Subsequently,the LrCOMT was ligated into pET-29a vector,and transferred into E.coli strain BL21 for heterologous expression.And the result demonstrated that the recombinant protein was induced by IPTG,and its molecular weight is about 40 kD,which is consistent with other COMTs from other plants.

Abstract:Based on the EST sequence encoded the glutathione S-transferase (GST),gene-specific primers were designed and used to obtain the full-length cDNA of a novel GST gene from Pyrus pyrifolia Nakai cv.Huobali in Yunnan Province with the method of rapid amplification of cDNA ends (RACE).This novel gene was named as PpGST.PpGST is 1 177 bp in length with an ORF of 696 bp,a 5′-untranslated region (UTR) of 130 bp,and a 3′-UTR of 351 bp,and the ORF encodes a predicted polypeptide of 231 amino acids.A phylogenetic analysis of the relationship of the newly identified PpGST with some known plant GSTs grouped the PpGST into the class of zeta GSTs.PpGST is abundantly expressed in pericarps of Huobali regardless received sunlight or not,but no expression was detected in the young leaves.The abundant expression of PpGST in fruits of P.pyrifolia hints that the PpGST may be involved in maintaining the redox balance and stress response during the pear fruit development.

Abstract:Comparison on the flower morphologic,pollen fertility alteration and utilization of male sterile line 160S in Brassica napus were conducted.The results showed that the temperature was the main factor to flower morphologic and pollen fertility alteration.Under male sterile conditions，the anthers of 160S were all smaller than that of normal ones and the pollens were completely male sterile.The percentage of pollen fertility,the self-pollenated pod-setting ratio and seed number per pod decreased along with temperature rising,and the fertility transformation direction was fertile,part fertile and then complete sterile.The results of test cross with varieties showed that all testers could restore the sterility of 160S at high restore ability.160S has high specific combining ability and heterosis,it can be used to produce hybrid seeds according to the two-line hybrid system model and is a prominent approach to utilizing heterosis in rapeseed.

Abstract:Two Arabidopsis mutants atmyb123 and atkor1 were identified from the T-DNA insertion knockout mutant lines SAIL_005260 and SAIL_2_G11,respectively,and then a double mutant atmyb123/atkor1 was established by crossing method.The two mutants are lacking expression for ATMYB123 and ATKOR1 genes,respectively,which two were found to be tightly related to root development in Arabidopsis thaliana.The results obtained here showed that lack of ATMYB123 gene in expression led to a slow growth of plant rosettes and a yellow skin of seeds in Arabidopsis,while lack of ATKOR1 gene in expression had no marked effects on these two factors.Any one of the two genes ATMYB123 and ATKOR1 knockout extremely repressed the root development in Arabidopsis,especially the knockout of ATKOR1 gene,the mutant atkor1 showed only one third of length of roots as compared to wild type (WT).Interestingly,the double mutant atmyb123/atkor1 exhibited the characteristics of the single mutant atmyb123 has in plant rosette morphology and seed skins but presented intermediated root length between the two single mutants.In addition,the growth trend of roots among the three mutants had no fundamental changes when the plants were cultivated under different pH,NaCl treatments and GA concentration conditions,which imply that these three factors were not concerned in the root shortening event induced by lack of any one of ATMYB123 or/and ATKOR1 proteins in A.thaliana.These results suggest that both ATMYB123 and ATKOR1 genes participate in the root development of Arabidopsis and a specific relationship in functions exist between the two proteins,ATMYB123 and ATKOR1.The transcription factor ATMYB123 might act as a major regulator of ATKOR1 protein for participating the control of root development in Arabidopsis.

Abstract:The genome data of Malus×domestica Borkh.‘Golden Delicious’ were downloaded from NCBI website,and 68 SSR primers i.e.four pairs from every chromosome were designed according to the SSR loci of genome and these primers were detected using 94 F₁ progenies of Pyrus ‘Huangguan’ (from Pyrus bretschneideri ‘Xuehuali’×Pyrus pyrifolia ‘Shinseiki’)×Pyrus bretschneideri ‘Laiyang Chili’,and the genetic diversity of this progeny was analyzed.The results were shown that:(1)the amounts of primer which could produce the expected product is 40 (58.82% of total primers),and 16 primers were shown polymorphic by PCR reaction.(2)Based on the results of amplification using the 16 polymorphic primers in the above progeny,the average number of alleles (Na) and the effective number of alleles (Ne) were 2.312 5 and 2.001 4,respectively.The mean observed heterozygosity (Ho),the mean expected heterozygosity (He) and the mean Shannon’s information index (I) were 0.548 3,0.490 5 and 0.746 2,respectively.16 polymorphic primers used for Pyrus L.were developed based on apple genome.The transferability of the SSR locus from genus Malus to Pyrus was also demonstrated in this paper.

Abstract:The differentce of protein subunits of 112 faba bean was analyzed by SDS-PAGE,in order to find out specific protein subunits.The result showed that:(1)the effective number of albumin and globulin alleles was 1.750 0±0.452 3,1.545 5±0.522 2,and the percentages of polymorphic subunit was 75.00%,54.55%,respectively.(2)It was difference between the albumin and globulin subunit numbers in different genotypes,containing 12 albumin subunits and 10 globulin subunits thereof the specific albumin subunits were 116 kD,96 kD,45 kD and specific globulin subunits are 58 kD,35 kD.42 varieties contain specific albumin subunit,21 varieties contain specific globulin subunit.(3)The lack subunits were 97 kD,63 kD of albumin and 97 kD,56 kD,47 kD of globulin.19 varieties were found with based albumin subunit lack and 21 varieties with globulin subunit lack.In summary,both albumin and globulin subunit numbers are different among different faba bean genotypes.Besides the based subunits,some have specific subunits and some have lack subunit.

Abstract:To understand the DNA methylation features of bamboo during its flowering process,we employed the techniques of HPLC (high performance liquid chromatography) and MSAP (methylation-sensitive amplified polymorphism) to detect the DNA methylation level of Bambusa multiplex (Gramineae,Bambusoideae),in a continuous period from its non-flowering to flowering stage.The results showed that DNA methylation rate of B.multiplex changed dynamically at different times.Furthermore,DNA methylation rates of flowering plant were significantly lower than those of non-flowering plant and non-flowering clumps.Total DNA methylation rates of 9.00% and 12.42% were detected in flowering and non-flowering plant,respectively,with full-methylation of 5.06% and 7.53%.Meanwhile,66.83% MSAP sites were identical both in flowering and non-flowering plants and 33.17% sites varied.Among those 33.17% variable sites,22.28% were demethylation and 1.98% were methylation in flowering plants,and other 8.91% variable sites gave a variance tendency between flowering and non-flowering plants.Therefore,demethylation and methylation happened simultaneously in flowering plants,with a much higher demethylation rate than that of methylation,thus exhibited a low methylation rate in flowering plants.

Abstract:Fluorescence microscope severed as a tool to study the pollen tube growth after crossing between different ploidy levels of Dianthus caryophyllus.Capsules setting rate,planting percent and index of compatibility were calculated.Chromosome counts were carried out in the hybrid progeny.The result shows that:pollen of ‘Zhenzhufen’ and ‘NH6’ germinated after 2 h and callose response was found in pollen tubes.Stylar cannal have callose response in 2 h after pollination.At 4 h,pollen tube tips were found in the style middle,callose response was found in pollen tubes.Some of the pollen tubes reached the base of the style after 6 h.Pollen tubes increased at the base of the style after 17 h,pollen tubes has reached the ovary,which has induced callose response.Fertilization of ovules was noticed from 17 h to 24 h after pollination,and the percentage of presumed fertilized ovules was very low.No seedlings were obtained in the ‘butterfly’（2n=60）בZhenzhufen’（2n=30）,three seedlings which were tetraploid were obtained in the ‘butterfly’（2n=60）בNH6’（2n=30）and revealed that the plants were possibly producing unreduced (diploid) gametes.

Abstract:Anther wall formation,microsporogenesis and the formation of male gametophyte in Swertia hickinii Burk.were reported for the first time in the present paper.(1)Anthers are etrasporangiate.Anther wall comprises epidermis,endothecium,one middle layer and tapetum and the development of anther wall is of the dicotyledonous type.The tapetum has dual origin and belongs to the glandular type.The endothecium and epidermis persist,but the cells in endothecium develop to become elongated and fibrously thickened.(2)The cytokinesis in the microsporocyte meiosis is simultaneous.Most of the microspore tetrads are tetrahedral and there are a few other types,such as isobilateral and “T”-shaped.Mature pollen grains are 3-celled and tricolporate.The common embryological characters of Swertia were generalized and compared to those of Gentiana,Crawfurdia and Tripterospermum.Whether the epidermis persists can be used to distinguish Swertia from Tripterospermum.

Abstract:Microsporogenesis and male gametophyte development of Caragana korshinskii Kom.were observed by traditional paraffin sections.The results showed that:(1)The anther had four chambers;the development of anther wall was the basic type,which consisted of epidermis,endothecium,middle layer and tapetum.The epidermis consists of one layer of cells and still existed as the anther matured.The endothecium developed banding-shaped thickenings when the anther matured.The young anther wall had 1~2 cell middle layer and degenerated when maturity of the anther,the tapetum was of glandular type.(2)Cytokinesis occurred simultaneously with meiosis in a microspore mother cell,and the tetrads were mostly tetrahedral while some were symmetrical.(3)The mature pollen grain was of binuclear type.With scanning electronic microscope (SEM),the mature pollen grains were observed,which appeared spherical in shape,and the exine was almost smooth.(4)Some of the tetrads generated by microspore mother cell had more than one nucleolus whose number changed from two to six,it may relate to the incomplete fuse of nucleolus in telophase two.These results suggested that microsporogenesis and male gametophyte development of C.korshinskii Kom.were normal.These observed results provided base materials for the reproduction biology and provided theoretical ground for exploitation and utilization of C.korshinskii Kom.

Abstract:The wild Lilium sulphureum,an important plant species that has medical and horticultural values,possesses bulbils as vegetative propagation.In this current work,we focused on the develpment of bulbil for L.sulphureum.Based on the morphological observation and anatomical study,the bulbil formation of L.sulphureum was identified.The bulbil was firstly appeared on the basal site of petiole in the axil.It originated from cells of sub-epidermal of petiole.The process of bulbil development could be divided into three stages:triggering phase,expanding phase,and ripeness phase.The maturated bulbils generally are spheral or ellipsoidal.The inner cells posses large mount of starch granule as storage assimilate.The germination rate of L.sulphureum bulbils is very high (>90%).This result showed that vegetative propagation of bulbils is an important way for ecological adaptation.

Abstract:The anatomical comparison of petioles from 38 species in Angelica sensu lato and allied taxa from China involved the shapes of cross section,smooth or undulate margin,the presence of cavity,the shape and number of collenchyma,and the number and arrangement of vascular bundle,which possessed rich diversity and intraspecies stability exhibiting taxonomic significance in those taxa.Based on these petiolar characters,four types of petioles could be recognized:Hollow-ring type (Ⅰ),Φ-type (Ⅱ),U/V-type (Ⅲ),Multi-ring type (Ⅳ).Moreover,the potential evolutionary trends of petiolar structure in this group were also suggested:solid petiole was derived;the Hollow-ring type seemed relatively primitive,the Φ-type was transitional,and both U/V-type and Multi-ring type were relatively developed.

Abstract:This paper made comparative study on the tissue structures of leaf,root and stem of the three Yunnan wild rice species and cultivated rice via hand-held slicing.It was found that (1) The tissue structures of leaf,root and stem between Yunnan wild rice and cultivated rice have significant differences,especially between the Jinghong Oryza meyeriana Baill.,Jinghong O.officinalis Griff.and cultivated rice.(2)In the leaf of main vein,Jinghong O.meyeriana Baill.has no gas chambers structure and the number of vascular bundles are the least with smaller vascular bundles areas,Jinghong O.officinalis Griff.and O.rufipogon Wall.have more than one vascular bundle and gas chambers structure,and the diameter of them are larger than cultivated rice,while there are two gas chambers in cultivated rice.(3)The structure of stem of Jinghong O.meyeriana Baill.is the smallest,and the vascular bundles is the fewest;The arrangement of vascular bundles is different in the stems wall from other rice.Jinghong O.officinalis Griff.and O.rufipogon Wall.have thicker stem and stem wall,more vascular bundles,and O.rufipogon Wall.stem exists aerenchyma structure.(4)In the root,O.rufipogon Wall.has more vessels,larger diameter of vessels and larger areas of stele than that of cultivated rice.O.rufipogon Wall.has exodermis grow casparian dots which have the function of casparian strips.This study lays the foundation of understanding the internal structures of wild rice,suggesting the relationship between the structures and the growth potential of Yunnan wild rice,the strong ability to absorb nutrients and resist diseases and insect pests,and exploring and utilizing the excellent genetic characteristics of wild rice.

Abstract:In this study,a susceptible cultivar from the cross of ‘Huacui’ and ‘Golden Delicious’ was used as our material,the interaction process of Alternaria mali and host leaves was examined with scanning electron microscope and transmission electron microscope in order to understand the dynamic changes of A.mali infects host.The results indicated that A.mali usually spread on the surface of host leaves not through stomatal apparatuses or other ways in the infection process.During the late stage,the chloroplast and plasma membrane of host leaves changed clearly,and many other cell organelles disintegrated,indicated that A.mali had excreted AM toxins acted on host cells,and the host cells started to decline.

Abstract:Four levels of drought stress were applied to 1-year-old potted seedlings of Sorbus folgneri to investigate the effects of drought on leaf anatomical structure and photosynthetic characteristics.The results showed that:(1)Drought stress had induced significant reduction in leaf thickness,caused higher palisade to spongy ratio in light and moderate,lower ratio in severe stress.(2)Drought depressed synthesis of photosynthetic pigments,reduced the tissue concentrations of chlorophylls and carotenoids,caused higher chlorophyll a to b and chlorophyll to carotenoids ratio.(3)The diurnal variation of net photosynthetic rate,transpiration rate,stomatal conductance decreased,but intercellular CO₂ density increased with drought stress increasing.The limitation of photosynthesis was mainly caused by non-stomatal factors.In summary,S.folgneri demonstrated to be drought-resistant,it could adjust its leaf anatomical structure and net photosynthetic capacity according to levels of drought stress to maintain survival and growth.

Abstract:Taking the dry powder of Datura stramonium L.leaves as the donor,the soybean seed as the acceptor,we studied the effects of allelochemical stress produced by aqueous leachate on seed germination,young root growth,root hair development,root-tip cell division of soybean by the indoor petri dish with filter paper.The result indicated:In the experiments concentrations scope,the soybean seed germination,young root growth and lateral root growth were inhibited with the dry powder aqueous leachate of D.stramonium L.leaves.And the young root gradually shortened and coarsened and appeared brown spot with the increased stress concentration.In addition,the high-frequency chromosome aberration and micronucleus were induced by allelochemical stress.But the root hair development and root-tip cell division showed a response that they were promoted when lower concentration and inhibited when higher concentration,to wit,the relative lower concentration significantly promoted root hair development and increased the mitotic index while inhibitory effects were observed at higher concentrations of allelochemical stress,the root-tip cell mitotic index obviously to drop.

Abstract:The growth,photosynthetic pigment content,activities of SOD,POD,and MDA content of three years old Tamarix chinensis under different salt-drought intercross stress were studied in order to clarify the effect of salt and drought stresses on the growth and physio-biochemical characteristics of T.chinensis seedling.The results showed that:（1）The influence of salt stress on growth was greater than drought stress,and the sensitivity of stem and leaf to salt and drought stress was higher than that of root system.The plant height,stem base diameter and dry matter of T.chinensis were all reduced with the salt stress intensified.（2）With the salt and drought stresses intensified,photosynthetic pigment content increased and then decreased,the major factor transformed from drought to salt stress.Under severe salt stress,the contents of chlorophyll a and b decreased significantly.（3）Under moderate drought stress,the activities of POD and SOD firstly increased and then decreased with the salt stress intensified.The oxidation resistance of SOD and POD weakened gradually with salt and drought stresses intensified.（4）Appropriate salt and drought stresses can weaken the content of MDA.However,the content of MDA was higher under severe drought and mild and moderate salt stress.The study indicated that T.chinensis can improve its stress resistance by adjusted self-growth and physiological and biochemical characteristics,and exhibited stronger drought and salt resistance.T.chinensis had tolerance to intercross salt-drought stress,and appropriate drought stress could increase the resistance of T.chinensis to salt stress.

Abstract:We choose Columbia wild type Arabidopsis thaliana as the experimental material,using He-Ne laser instrument(632.8 nm,5 mW/mm ²) to handle the seeds,then gather statistics about germination potential,germination percentage,root length,the contents of chlorophyll,soluble sugar,soluble protein and MDA.The results showed that,short period of He-Ne laser radiation could promote the growth of A.thaliana.It has an obvious influence when the time reaches 4 minutes,and the germination potential,germination percentage,root length,the contents of chlorophyll,soluble sugar,soluble protein reach the highest.However,the content of MDA reach the lowest.When the time is beyond 4 minutes,the promotion influence become less,and then it inhibited with longer radiation time.All of these show that suitable He-Ne laser dosage radiation could promote the seeds germination and seedling growth of A.thaliana and protect the cell membrane from being injured.

Abstract:Reaumuria trigyna seedings were treated with NaCl of different concentrations (0,100,200,300 and 400 mmol/L) for 30 days.A study about its growth,physiological and biochemical mechanisms in different NaCl concentrations showed:(1)Under 100 and 200 mmol/L NaCl stress,the fresh weight and dry weight increased significantly than that of control.Growth of R.trigyna was inhibited with increasing NaCl concentrations,showing that the suitable NaCl concentration of R.trigyna seedlings growth was 200 mmol/L.(2)P_n,T_r and G_s decreased and C_i increased,showing decrease of photosynthesis rate causing reduction of utilization rate and intracellular CO₂ accumulation which was higher when NaCl concentration was higher.(3)Osmotic adjustments of R.trigyna mainly relied on Na⁺ and Cl^-,soluble sugar,free amino acid,proline and soluble protein.This study discovered that R.trigyna could improve its salt tolerance ability by adjusting Na⁺ and Cl^- content and osmosis regulating substances in leaves.