August 1997, Volume 39 Issue 8


          Research Articles
A New Approach for Gene Transfer into Citrus
Author: Li Wei, Chen Liang, Cai De-tian and Yang Guan-pin
Journal of Integrative Plant Biology 1997 39(8)
    Agrobacterium-mediated in vivo transformation of lateral buds of citrus plants has been developed and been combined with in vitro cloning. This new approach has ingeniously detoured plant regeneration and the long juvenile stage of regenerated plants from protoplasts, which limits gene transformation of most fruit trees including citrus. It makes the elimination of remaining bacteria and the selection of transformants be easy and efficient by using the natural growth of buds. In addition, this approach has combined the cloning of transfonnants in order to increase population rapidly. In demonstrating experiment, 21 out of 112 treated buds of Shatian pummelo ( Citrus grandIs Osb. ) developed into primary branches which gave 76 secondary buds. 4 in 76 secondary buds (5.26%) were proved to be positive by GUS activity assay. Among 14 cluster buds differentiated from the in vitro culture of 4 positive buds, 4 showing highest GUS activity and being positive in Southern hybridization were regenerated into tree plants. This approach could be used as a simple and effecient method for the transformation of a wide range of fruit tree species.
Abstract (Browse 1764)  |  Full Text PDF       
In Vitro Divisions of Unfertilized Central Cells and Other Embryo Sac Cells in Nicotiana tabacum var macrophylla
Author: Fu Ying, Sun Meng-xiang, Yang Hong-yuan and Zhou Chang
Journal of Integrative Plant Biology 1997 39(8)
    The embryo sacs and female cells could be isolated from the unfertilized ovules of Nicotiana tabacum L. var. macrophylla which were treated in a solution containing 1.5 % cellulase R- 1O, 1% macerozyme R-10, 10% mannitol, 10 mmol/L CaCI:, pH 5.8 for 3 h followed by given slight pressure with a micropipette. The central cells could be kept viable for 10 h and the egg cells for 3 h in 10% mannital. Sometimes, the in situ fusion products of egg cell and synergid protoplasts could be obtained and kept viable for at least 5 h. The high concentration (20 mg/L) of 2, 4-D was used in enzyme solution to induce the division of the unfertilized central cells and other megagametophytic cells in subsequent culture. Treatment of 2,4-D together with enzymatic maceration of ovules was proved to be better than its direct treatment of isolated embryo sac or its component cells. Isolated embryo sacs were cultured in microchambers (Millicell-CM PICM 012 50 MILLIPORE) feeded with divided mesophyll protoplasts of Nicotiana rustica L. The medium was KMSp medium supple- mented with 1% glucose, 0.1 mol/L mannitol, 0.1 mol/L sorbitol, 0.25 mol/L sucrose, 1 mg/L BA, 6% to 10% coconut water, and 0.15% low gelling agarose. Division of central cells, antipodal cells and the in situ fusion products of egg cell and synergid protoplasts were induced. The unfertilized central cell was for the first time to be induced in vitro to develop into small cell clusters.
Abstract (Browse 1708)  |  Full Text PDF       
Osmotic Stress Decreases the Activity of ATPase Associated with the Root Cap Plasmodesmata in Zea mays
Author: Miao Long, Wang Xue-chen and Zhang Wei-cheng
Journal of Integrative Plant Biology 1997 39(8)
    With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L. ) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the departed periplasm and stretching towards cell wall could be observed even after plasmolysis. By observing the precipitation of ATPase activity product (lead phosphate) at plasma membrane and plasmodesmata, it was found that the fine strands were plasma membrane-lined channels surrounding the cytoplasm and that they still firmly connected to the plasmodesmata during plasmolysis. Compared with the control (unstressed), a sharp decrease of ATPase activity in the plasmodesmata of the stressed cells was observed. Inhibition of energy metabolism in these limited locales would affect the physiological activity, maybe including the regulation of permeability and the change of size exclusion limit (SEL) of plasmodesmata.
Abstract (Browse 1853)  |  Full Text PDF       
GUS Activities Under Control of HRGP Promoter of Carrot
Author: Yin Shu-ping, Guo Yang, Ma Mi, Hu Yuan-lei, Zhao Wei-ping and Lin Zhong-ping
Journal of Integrative Plant Biology 1997 39(8)
    Hydroxyproline rich glycoprotein (HRGP) promoter has been cloned from genomic clone of carrot ( Daucus carom var. sativa DC. ) by PCR procedure. The construction with fusion gene of carrot HRGP/GUS in binary vector were used for the genetic transformation by Agrobacter/um mediated procedure and the transgenie tobaccoes with this transgene were obtained. The GUS activities in various organs have been studied by histochemical examination. Tissue specific activities of this HRGP promoter were demonstrated. GUS activities were expressed in the apical meristematic zones of roots, inner phloem of stems, inner and outer phloem of petioles. The highest levels of GUS activity were found in tissues where axillary bud was initiated. The expression of GUS activities under the control of carrot HRGP promoter could also be enhanced by treatment with the inducing factors such as salicylic acid, CaC12 and NaC1.
Abstract (Browse 1750)  |  Full Text PDF       
Isoflavones from Astragalus membranaceus
Author: Song Chun-qing, Zheng Zhi-ren, Liu Di, Hu Zhi-bi and Sheng Wan-yun
Journal of Integrative Plant Biology 1997 39(8)
    Two new isoflavones (8, 3'-dihydroxy-7,4'-dimethoxyisoflavone, odomtin-7-O--D-glu-copyranoside) and four known isoflavones (formononetin, 7,3' -dihydroxy-8,4'-dimethoxyisoflavone, ealyeosin, ealycosin-7-O--D-glucopyranoside) were isolated from the roots of Astragalus merebtm (Fiseh.) Bunge. Their structures were established by spectral analysis.
Abstract (Browse 1933)  |  Full Text PDF       
The Relationship Between Pollen Flux and Meteorological Elements and Its Prospect as a Technique of Crop-Output Prevision of Wheat and Maize
Author: Huang Ci-xuan, Liang Yu-lian, Chen Zhi-qing and Cheng Zhi-gang
Journal of Integrative Plant Biology 1997 39(8)
    The crop output has been successfully forecasted for more than ten years on the basis of pollen flux abroad. The annual crop output depends upon meteorological elements (temperature, wind velocity, precipitation, etc. ). These elements have influence on the pollination intensity (airborne pollen density, number of pollen grains' m- 3 air), which reflects directly the crop output. During 1992 to 1994, the authors had collected and measured the airborne pollen with Cour' s sampler, and recorded the major meteorological data. From statistical analysis, the authors have obtained data on the relationship between the outputs of wheat and maize and their pollen deusity during the maximum pollination period. Thus formulating a prehminary model for crop-output prevision of wheat and maize has been attempted.
Abstract (Browse 1800)  |  Full Text PDF       
Genetic Differentiation of Populus cathayana
Author: Li Kuan-yu, Huang Min-ren, Yang Zi-xiang, Wang Ming-xiu and Huang Feng-hou
Journal of Integrative Plant Biology 1997 39(8)
    169 clones chosen from 30 localities of Populus cathayana Rehd. covering ten provinces were colleced for analysis of DNA polymorphisms between populations and within population by RAID. The result showed that the degree of DNA polymorphism between populations was much larger than that within population, and the degree was greater between eastern and western population than that between the neighbor populations. The UPGMA and Neighbor-Joining cluster analysis based on net genetic distance were used to investigate phylogenetic relationship among populations. They beth showed apparent genetic differentiation between eastern and western population. The dendrogram had correspondence with geographic distribution region of P. cathayana. The western revealed much more polymorphisms than the eastern population. Therefore, the western populations had more important role in the origin and evolution of P. cathayana. According to the relationship between diversity center and origin centerof species, it is possible that P. cathayana originated from the northwestem China and then spreaded to the eastern region forming modem distribution region.
Abstract (Browse 1823)  |  Full Text PDF       
Responses of Rice Seedlings in Different Drought Tolerant Cultivars to Oxidative Stress
Author: Guo Zhen-fei, Lu Shao-yun, Li Bao-sheng, Li Ming-qi and Li Yong-chao
Journal of Integrative Plant Biology 1997 39(8)
    Rice ( Oryza sativa L. ) of drought-tolerant cuhivar "Xiangzhongxian No. 2" showed much lower O2- production rate under drought condition than that of drought-sensitive cultivar "Xi- angzaoxian No. 12". Drought damages on both rice cultivars were accelerated by paraquat (0.01 mmol/L) and H2O2(10 mmol/L); but the drought-tolerant cultivar had greater resistance to these damages than did the drought-sensitive cuhivar. The results also showed that in rice of droughttolerant cultivar, the increase in activities of. superoxide dismutase and peroxidase were much higher than that in drought-sensitive cultivar after treatment of paraquat, and the increase in activity of catalase was more significantly after treatment of H2O2 than that in drought-sensitive cultivar. These results suggest that the drought-tolerant cultivar had greater resistance to oxidative stresses than did the drought-sensitive cultivar.
Abstract (Browse 1891)  |  Full Text PDF       
Preparation of Gui Culture Medium and Its Application to Wheat Anther Culture
Author: Hai Yan, He Xian-chang, Huang Bing-yan and Wang Jin-lan
Journal of Integrative Plant Biology 1997 39(8)
    Gui culture medium has been developed by adjusting the concentration of macro- and micro- elements, based on the studies of various media applying to wheat ( Triticum aestivum L. ) anther culture breeding for many years. An appropriate ration among NH4N03, KN03, MgSO4 and MnSO4 in the medium was selected by orthogonal design L16 (45). For comparison of different media namely C17, W14 and N6 medium, the average frequencies of callus induction were increased by 30.31%, 50.66% and 57.96% respectively when Gui medium was used. And the Gui medium showed all its advantages in various crosses. The frequencies of wheat anther callus induction showed an increase of 54.25 % to 64.07 % and the callus development was also promoted when adding 0.9 mg/L rare earth addition to the Gui medium.
Abstract (Browse 2109)  |  Full Text PDF       
Effect of Host Plants and Inoculum Forms on the Inoculum Potential of Arbuscular Mycor rhizal Fungi
Author: Yin Xi-sheng, Liu Run-jin and Sun Xian-ming
Journal of Integrative Plant Biology 1997 39(8)
    The effects of inoculum forms (single-spore, multi-spores, or colonized root pieces) and host plants (Nicotiana tabacum L., Sorghum sudanense(Piper) Stapf, and Trifolium repens L. ) on the development and inoculum potential (IP) of the arbuscular mycorrhizal fungi (AMF): Glomus macrocarpum Tul & Tul, Glomus mosseae (Nicol & Gerd. ) Gerdemann & Trappe, Glomus versiforme (Karsten) Berch, and Sclerocystis sinuosa Gerdemann & Bakhi cultured in pots were investigated. The lag phase of treatment with 50 spores or 0.5 g (fresh weight) of colonized root pieces was 4 weeks, much shorter than that of the treatment with 1 spore (8 weeks); the value of IP(VIP) and percentage of root colonization(PRC) of the former were greater than those of the latter. Only on the early stages of colonization was there difference between the 50 spores and the 0.5 g (fresh weight) of colonized root piece inoculation treatments. The IP per plant inoculated with 0. 5 g (fresh weight) of colonized mot pieces of AMF was greater than that of the other two treatments except G. vers/forme on Nicotiana tabacum, while the PRC of the plants inoculated with 50 spores and 0. 5 g (fresh weight) of colonized root pieces of AMF was higher than that of the 1 spore inoculation after 10 weeks. The VIP of AMF on Trifolium repens was significantly higher than that on the other two hosts. The VIP of G. mosseae, G. versiforme, and S. sinuosa was respectively greater than that of G. macrocarpum. This suggested that different species of AMF produced different VIP of the inoculum. Nicotiaha tabacum was much better than the other host plants which used to be inoculated with single spore, and to produce inocula of AMF.
Abstract (Browse 1923)  |  Full Text PDF       
Standardization of Electrophoregrams for Classification and Identification of Wheat Cultivars
Author: Sun Zhi-liang, Jiang Jia-hui, Liang Yong-ping, Sun Mei-juan, Tan Ye-jie and Peng Shou-hua
Journal of Integrative Plant Biology 1997 39(8)
    Electrophoregrams of four wheat ( Triticum aestivum L. ) cultivars were evaluated for band deviations and control of methods using 9% acrylamide gels in a glycine acetic acid buffer at pH 3.10. The regular patterns of band deviation changed with its migration distance which revealed that the farther the band moved, the greater its standard deviation and range. Comparing with band distance of 2.05 cm, the band at 9.25 cm had a three fold increased standard deviation of 0.033 2 and the range increase from 1.5 mm to 4.5 mm. No biotypes of electrophoregrams was observed in "Yannong 15" ,"Jinan 13" and "Laizhou 953". The results demonstrated that "Laizhou 953" and "Jinan 13" were better reference cuhivars than "Marquis". Four marker bands 2.05 cm,3.15 cm, 6.05 cm and 9.25 cm were chosen based on the assessment of the electrophoregrams. In accordance with the standard averages of these markers and their actural migrating distances in a nm a calibration coefficient and calibrated migrating distance could be computed for each band on an elec- trophoregram. Thus a two third reduction of the standard deviation could be expected.
Abstract (Browse 1809)  |  Full Text PDF       
Plant Regeneration from Asymmetric Somatic Hybrids of Oryza sativa and Panicum maximum
Author: Xin Hua-wei, Sun Jing-san, Yan Qiu-sheng and Zhang Xue-qin
Journal of Integrative Plant Biology 1997 39(8)
    Asymmetric fusion of rice ( Oryza sativa L. ) protoplasts with protoplasts of apomictic Panicum maximum Jacq. by means of polyethylene glycol (PEG) method has been resulted to regenerate plants after selection and culture of the fused cells and that could be transplanted alive into soil. Before fusion, rice protoplasts were treated with 2.5 mmol/L iodoacetamide (IOA) at room temperature (22 to 25 ) for 15 min, while protoplasts of P. maximain were irradiated with 60 Kr soft X-ray. Dramatic variation was found on the flower organ morphology and stmcture,as well as re-production characteristics, etc. in the 28 flowering plants obtained. All of the plants showed abnormally increased number of anthers, ranging from 7 to 13. In some plants there were 2 to 3 ovules in a single ovary. In two plants there appeared to be more than 2 embryo sacs in a single ovule. Female and male fertility decreased greafiy and even vanished thoroughly. Only 5 regenerated plants produced seeds with low frequency. I-KI stainable pollen ranged from 0 to 68%. Cytoembryological observance showed that the non-seed-setting plants were female sterile,i.e. ,no normal embryo sac was differentiated in these plants.
Abstract (Browse 1713)  |  Full Text PDF       
Studies on Special Proteins and Polypeptides of Sperm Cell-Rich Fraction in Orychophragmus violaceus
Author: Dong Chang-jiang, Zheng Chang-wen, Li Bing-chen, Liu Zhi-hui and Huang Yu-xiang
Journal of Integrative Plant Biology 1997 39(8)
    d sperm cell-rich) were isolated from disrupted pollen of Orychophragmus violaceus (L.) O.F. Schulz with 10 lan nylon filter and sucrose gradient centrifugation. Proteins were extracted using a phenol procedure and polypeptide patterns were analysed with two-dimensional polyacrylamide gel electrophoresis. The sperm cell-ri& fraction had 249 spots and the sample fraction contained 204 spots. The interleaf of sample and 30% sucrose fraction revealed 190 spots, the 30% sucrose fraction 244 spots and the interleaf of 30% and 40% sucrose fraction 167 spots. Compared the spots of the sperm cell-ri& fraction with those of the other four fractions, more than 18 spots were unique to the sperm cell-rich fraction and most of the unique spots were above 60 kD.
Abstract (Browse 1628)  |  Full Text PDF       
A Novel Floral Mutant of Petunia
Author: Guo Feng-li and Meng Fan-jing
Journal of Integrative Plant Biology 1997 39(8)
    A new kind of petunia ( Petunia hybrida L. ) floral mutant, designated as e fficient (eff), was characterized as having increased ntanber of organs in all floral whorls compared with the wild type. The number of carpel was steadily increased up to 3 to 4. The pistil had a 3- or 4-lobbed stigma. Meanwhile, petaloid structures developed on top of the stamens, but pollen formation and anther dehiscence were not affected. After fertilization, the mutant flower formed a larger fruit consisted of more than two loculi. Scanning electron microscopic observations revealed that the increased organ number and petaloid structure occurred at primodia stage in the mutant. This eft mutation acted as a single Mendelian trait in genetic crosses. The phenotype of the mutant flower are advantageous to sexual reproduction process of plant.
Abstract (Browse 1703)  |  Full Text PDF       
Autoinhibition in Polar Transport of Indoleacetic Acid
Author: Li Chun-jian
Journal of Integrative Plant Biology 1997 39(8)
    Autoinhibition in polar transport of 3H-IAA from donor to receiver was demonstrated in pea ( Pisum sativum L. ) and mung bean ( Phaseolus radiatus L. ) using either classic "donor-receiver system" where unlabelled IAA was present in the receiver or using "Y" form explants where tmlabelled IAA in lanolin was applied to the cut surface of a separate side of the explants. Application of unlabelled IAA resulted in accmnulation and high rate of metabolism of SH-IAA in the labelled tissues, and these were further stimulated by prolonging the experimental period.
Abstract (Browse 1727)  |  Full Text PDF       
Sequence Analysis of Rice Lipid Transfer Protein cDNA and the Structural Characteristics of Corresponding Peptide
Author: Zhan Shu-xuan, Ge Xiao-chun, Li Geng, Sun Chong-rong and Cao Kai-ming
Journal of Integrative Plant Biology 1997 39(8)
    A eDNA done containing a putative coding region for lipid transfer protein (LTP), designated as pFDRSC144, was isolated from a rice ( Oryza sativa L. ssp. indica )culfivar "Guanglu' ai 4", etiolated shoot cDNA library, 840 bp nucleotides of cDNA was sequenced and it contained an open reading frame of 354 nucleofides encoding 118 amino acids. The deduced peptide included a signal peptide of 26 aa, and showed high homology with LTPs from rice and other plants. The hydropathy profile revealed that the putative peptide chain contained several hydrophobic regions. The secondary structure analysis by the method of Gamier showed that this peptide chain of mature protein could mainly form -pleated sheet structure. It has been therefore tentatively pmposed that LTP peptide might form an open tube-like structure which would be useful for lipid transfer. In addition, the result of Southern blot analysis revealed the existence of'an LTP gene family in the rice genome.
Abstract (Browse 1732)  |  Full Text PDF       
Microdissection and Recovery of 1R Chromosome in Rye ( Secale cereale )
Author: Cui Li-hua, Hu Zan-min, Wang Lan-lan, Li Liang-cai and Chen Zheng-hua
Journal of Integrative Plant Biology 1997 39(8)
    The technique of mieromanipulaion has been used to establish a system of single chromosome mierodissection. According to the standard karyotype of rye ( Secale cereale L. ), 1R chromosome carrying disease-resistant gene has been identified, microdissected and transferred into Ep-pendorf tube using mieromanipulator. The results showed that IR chromosome in cells pretreated with -bromonaphthalene could be identified quickly and the efficiency of microdissection was greatly improved when the technique of wall degradation and hypotonic treatment was applied.
Abstract (Browse 1795)  |  Full Text PDF       
Actin is a Constituent of the Nuclear Matrix and Chromosome Scaffold in Physarum poly cephalum
Author: Zeng Xian-lu, Jiao Ming-da, Wang Xiao-guang and Hao Shui
Journal of Integrative Plant Biology 1997 39(8)
    Nuclear matrices and chromosome scaffolds were obtained by digesting and extracting, respectively with DNase and 2 mol/L NaC1, the nuclei and chromosomes isolated from the plasmedia of Physarum polycephalum Schw. The results of the indirect immunofluorescence of tacit antiactin antibody as immunomarker indicated that the nuclear matrices and chromosome scaffolds both had positive reaction with the antibody. The results of the indirect immunodotting experiment further verified the presence of actin antibody in their constituent. Protein A-colloidal gold immunoelectron microscopy technique revealed that gold particles were distributed in the interphase nuclei and metaphase chromosomes. The above results showed that actin is a constituent of the nuclear matrix and chromosome scaffold of P. polycephalum.
Abstract (Browse 1740)  |  Full Text PDF       
Actin is Immunolocalized in the Nuclei and Chromosomes of Vicia faba
Author: Wan Li-hong and Xing Miao
Journal of Integrative Plant Biology 1997 39(8)
    Meristematic cells of Vicia faba L. were labeled with rabbit anti-actin antibody and FITC-conjugated goat anti-rabbit lgG antibody and observed with fluorescence microscopy. Both the nuclei and chromosomes sent forth distinctive fluorescence, indicating that actin is present in the nuclei and chromosomes. Sections were reacted with the anti-actin antibody and protein A-colloidal gold and observed with transmission electron microscopy. Gold particles were found over the whole nuclei, and a lot of particles were concentrated in condensed chromatin areas and nucleoli, confirming the observations with the fluorescence microscopy. V. faba nuclei and chromosomes were treated with DNase and 2 mol/L NaC1, and DNA and histone-depleted nuclei and chromosomes were obtained. Indirect immunofluorescence tests showed that the DNA and histone-depleted nuclei and chromosomes reacted positively with the anti-actin antibody. These results demonstrated that actin exists not only in intact nuclei and chromosomes but also in DNA and histone-depleted nuclei and chromosomes of V. faba. In addition, the authors' results indicate that tropomyosin is present in the nuclei and chromosomes of V. faba. Presence of actin in nuclei and chromosomes as well as in DNA and histone-depleted nuclei and chromosomes of higher plants is discussed.
Abstract (Browse 1860)  |  Full Text PDF       


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