May 1998, Volume 40 Issue 5


          Research Articles
Distribution of Gibberellins A7 and A4 in Tobacco Proembryos Using Immunoelectron Microscopy
Author: CHEN Yi-Feng, LIANG Shi-Ping, YANG Hong-Yuan, ZHENG Zhi-Fu and ZHOU Xie
Journal of Integrative Plant Biology 1998 40(5)
    The ovules of Nicotiana tabacum var. macrophylla 8 days after pollination were fixed successively with 2% EDC (1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide) and a mixture solution of paraformaldehyde and glutaraldehyde, then slightly postfixed in 0.5 % osmium solution. UI- trathin sections of 6-, 9- or 12- celled proembryos embedded in Epon 812 resin were stained in an anti-GA MAb and sheep anti-mouse IgG-colloidal gold (10 nm). This MAb specifically recognizes methyl esters of GA7 and GAn. Therefore, it could be used as a probe to localize GA7 and GAn in cells after EDC fixation. The 12-celled proembryo is composed of a 9-celled embryo and a 3-celled suspensor. Wide distribution of GA7 and GAn was observed in all proembryo cells and most organelles at subeellular level, including walls, plasmodesmata, plasma membrane, cytoplasmic matrix, mitochondria, plastids, vacuoles, endoplasmic reticulum and nuclei. Clusters of gold granules were found in nuclear envelope, nucleomatrix, nucleolus, chromosome, cytoplamic matrix. In a region composed of cytoplasmic matrix, a vacuole and a mitochondrium, such concentrated gold granules were particularly obviously observed. There appeared a gradient distribution of GA7 and Gan from embryo cells decreasingly to suspensor cells. GA7 and GAn could be translocated via intercellular walls, plasmodesmata and vesicles between embryo and suspensor. 1he authors suspect the direction of GA translocation in proembryo may be from suspensor to embryo. To author's knowledge, this is the first report to indicate subcellular and gradient distributions of bioactive gibberellins in plant proembryos.
Abstract (Browse 1860)  |  Full Text PDF       
Acceptance and Analysis of Plant Science Projects Supported by the National Natural Science Foundation of China in 1997
Author: ZHU Da Bao, LIU JiaXi and WANG Yun
Journal of Integrative Plant Biology 1998 40(5)
Abstract (Browse 1862)  |  Full Text PDF       
Correlation Between Appearance of Embryogenic Cells and the IAA Levels in Rice Somatic Cell Culture
Author: CHEN Yi-Feng, ZHOU Xie, TANG Ri-Sheng, ZHANG Jin-Yu and MEI Chuan-Sheng
Journal of Integrative Plant Biology 1998 40(5)
    Changes of endogenous IAA level and IAA action in cultured rice ( Oryza sativa L. ) somatic cells during the period from 7th to 15th day which was the transition from somatic to embryogenic cells were observed. The study was carded out in three experimental systems viz. mature caryopsis and young panicles (2 ~ 5 mm long) of rice cv. "Guangluai 4" under normal osmosis (3% sucrose), mature caryopses from rice cv. "Yanjing 2" or "Guangluai 4" under normal and higher osmosis (5% sucrose or 2.5 % sorbitol). During this period, endogenous IAA contents were greatly increased in young-panicle calli under normal osmosis and mature-caryoptic calli under higher osmosis but decreased in mature-caryoptic calli under normal osmosis. Exogenous IAA could induce the appearance of embryogenic cell from nonembryogenic callus at a lower frequency. And 2,3,5-tri-iodobenzoic acid could increase the frequency of embryogenic cell induction. From these results it could be concluded that accumulation of higher IAA level in the cultured rice cells was essential for induction of embryogenic cell appearance. Since 2,4-D was involved in all induction medium with the same concentration but exerted different effects on embryogenic cell induction, it was suggested that it might act through mediating the endogenous IAA metabolism.
Abstract (Browse 2084)  |  Full Text PDF       
Genetic Transformation of Cucumis sativus by Agrobacterium rhizogenes
Author: SHI He-Ping, LI Ling and PAN Rui-Chi
Journal of Integrative Plant Biology 1998 40(5)
    Hairy roots were obtained in vitro 10 days after inoculation of cucumber ( Cucumis sativus L. ) cotyledon explants with the strains of Agrobacterium rhiwgenes R1000 and R1601. The frequency of the cotyledon explants transformed by R1000 and R1601 was up to 87.5% and 88.9%, respectively. All hairy roots induced by the strains of R1000 and R1601 grew rapidly on solid hormone-free MS medium. The roots incited by A. rhizogenes R1000 could be divided into three phenotypes. The roots of phenotype were similar to the normal ones, but had more numerous lateral roots. Roots of phenotype m were much stouter and shorter, they elongated very slowly and were more highly branched than roots of phenotype . Roots of phenotype were of intermediate in appearance. However, the roots incited by A. rhizogenes R1601 appeared similar to phenotype roots incited by A. rhizogenes R1000. Transformation was confirmed by opine detection.
Abstract (Browse 1949)  |  Full Text PDF       
Studies on the Characteristic of Element Contents in the Dominant Plant Species of the Threegorges Region in China
Author: HE Jin-Sheng, CHEN Wei-Lie and WANG Qi-Bing
Journal of Integrative Plant Biology 1998 40(5)
    The characteristics of the contents of 20 elements (Al, Fe, Mn, Ca, Mg, K, S, Si, P, Cd, Cu, Zn, Ti, Ni, Sr, Mo, Na, B, Cr, V) in 16 plant species collected from the Three Gorges Region in China were investigated. The average contents of Ca, K and Mg were higher than 1 000 gg-1, that of Al, P, Si, Fe, S and Mn ranged between 1001 000 gg-1 and Ti, Cu, Ni, Cr, Mo, Cd and V were less than 10 gg- 1. The level of Na content was less than that of the reported. The main character of the element contents was of the Ca> K type. The contents of P, S, Ca and K in different plant samples showed a normal distribution pattern, while Al and Mn showed a elements lognormal distribution pattern. Plant species differed greatly in the element contents. On analyzing the coefficient of variation (C. V., % ), Al, Mn, Mg, Ni, Sr and Fe had higher C.V., while the C.V. of K, S, P, Cr, Cd and Cu was less than 60%, and Cu had the lowest C.V. The correlations between Al and Fe, Al and Ti, Al and Cr, A1 and V, Cd and Sr, Cd and Mo, Fe and V, Zn and Cr, Ni and Sr, Mg and Ni, Mo and Sr, Ca and Sr, Cr and Mo, Na and Mg, Na and P, P and S were statistically significant in different plant species. The classification of the 16 plant species and 20 dements by two-way indicator species analysis (TWINSPAN) method may suggest the difference in dement contents of the different plant species.
Abstract (Browse 1972)  |  Full Text PDF       
Establishment of Hairy Root Cultures of Astragalus membranaceus and the Extrinsic Factors Affecting Their Growt
Author: HU Zhi-Bi, ZHENG Zhi-Ren, LI Xing-Ping and LIU Di
Journal of Integrative Plant Biology 1998 40(5)
    Hairy root cultures were established from the leaf explant of Astragalus membranaceus (Fisch.) Bge. infected with Agrobacterium rhizogenes and factors that affect their growth were investigated. It was found that the hairy roots grew best in a phytohonnone-free MS medium at pH 6, in which the most optimal concentration of sucrose was 3 % and NH4NO3 was omitted because of its inhibitory effect to the hairy roots. Light was also inhibitory to them but the effect of low temperature was not apparent. Moreover, there were more crude saponin and soluble polysuccharide contents in the hairy roots than in the natural dry roots. Finally, a discussion on the application and the potenrial of the hairy root technique in the production of the plant roots as an effection Chinese traditional medicine was also involved.
Abstract (Browse 2031)  |  Full Text PDF       
Ultrastructural Studies on the Secretory Cavity Development and Essential Oil Accumulation in the Fruits of Evodia rutaecarpa
Author: LIU Wen-Zhe,ZHANG Hong and HU Zheng-Hai
Journal of Integrative Plant Biology 1998 40(5)
    The secretory cavity in fruits of Evodia rutaecarpa (Juss.) Benth. developed schizogenously through a separation of the walls of the central initial cells. Electron micrographs revealed that in the early stages of cavity development there was an apparent increase in the number and volume of the plastids in which esmiophilic droplets and tubular dements were observed. This suggested that the essential oils might be synthesized in the plastids.. The essential oils were then transport ed through the plastid membrane to the surrounding endoplasmic reticulum or into the vacuoles, becoming vesicles approaching the plasmalemma, and finally releasing their contents into the oil chamber by plasmalemma invaginations.
Abstract (Browse 1962)  |  Full Text PDF       
The Puff-like Structure: An Intranuclear Structure for Synthesis of Ribonucleic Acid Polymerase Transcripts as Revealed by the 5-bromouridine-5-triphosphate Labelling and Immunoelectron Microscopy
Author: HU Bo and XING Miao
Journal of Integrative Plant Biology 1998 40(5)
    The puff-like structure (PIS) is an intranuclear structure often observed in cells of higher plants. Several hypotheses regarding the nature of the structure have been put forward while the opinions are still controversial. By using the 5-bromouridine-5'-triphosphate (BrUTP) labeling technique which was developed in early 90s and has been utilized to localize nascem RNA polymerase (RP ) transcripts, in combination with immunoelectron microscopy. The authors studied PISs in the nuclei of the meristematic cells of Allium cepa L. A large number of gold particles were observed over the PISs of the specimens labelled with BrUTP and anti-BrdU antibody, indicating that RP transcripts were being synthesized actively in the structure. When the specimens were treated with a-amanitin ( 10 g/L, 2 h) which inhibits the activity of RP , and followed by treatment with BrUTP and anti-BrdU antibody, gold particles in the PLSs of the specimens were decreased dramatically and the gold density of the structure dropped strikingly from 66.65/m2 to 1.77/m2, confirming that the gold particles in the structure represented the RP H transcripts. It further demonstrated that the PIS was an intranuclear structure essential for the synthesis of RP transcripts.
Abstract (Browse 1817)  |  Full Text PDF       
Ultrastructural and Cytochemical Study on Mature Pollen of Pinus tabulaeformis
Author: YUAN Zong-Fei and HU Shi-Yi
Journal of Integrative Plant Biology 1998 40(5)
    The pollen of Pinus tabulaeformis Cart. comprised two prothallial cells, a generative cell and a tube cell which degenerated at pollen maturation. The generative cell had its own cell wall, seperating from the intine of pollen, but with its side wall attached to the infine. Cytoplasmic channels were present on the side of the generative cell wall, which faced to the tube cell cytoplasm. The generative cell differed conspicuously from the tube cell. The main differences include: ( 1 ) The chromatin in the generative cell nucleus was condensed, but was dispersed and had numerous nueleare pores in the tube cell nucleus; (2)There was no microbody in the generative cell but many microbodies were present in the tube cell cytoplasm; (3)More inclusions were present in the tube cell than in the generative cell. Both the generative cell and the tube cells contained lipid bodies and amyloplasts in the cytoplasm, but there were more amyloplasts in the former. The tube cell also contained a few proteins which was absent in the generative cell. In addition, there were numerous mitochondria, polyribosomes, and a few endoplasmic reticulums and dictyosomes in the generative and tube cells. DAPI staining demonstrated numerous cytoplasmic DNA in both generative cell and tube cell. The mode of cytoplasmic inheritance, and the composition, structure and the nature of the pollen wall of P. tabulaefonnis are also discussed in this paper.
Abstract (Browse 1926)  |  Full Text PDF       
Retardation of Leaf Senescence by Triadimefon Treatment in Mung Bean Seedling
Author: GUO Zhen-Fei, LU Shao-Yun, LI Bao-Sheng and LI Ming-Qi
Journal of Integrative Plant Biology 1998 40(5)
    Treatment of triadimefon on detached leaves of mung bean (Phaseolus radiatus L. ) seedlings increased the levels of chlorophyll and soluble proteins. Declined activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate-peroxidase (AsA-POD) and contents of ascorbate (ASA) and glutathione (GSH) were observed during the senescence of detached young leaves. Triadimefon at concentration of 20 mg/L promoted the activities of POD, AsA-POD and levels of AsA and GSH, but had no effect on the activities of SOD and CAT. On the other hand, the levels of malondialdehyde (MDA) were increased and the increase of which was markedly negative correlated with the activities of POD, AsA-POD and with the contents of AsA and GSH during the senescence of leaves. MDA contents were decreased by triadimefon treatment. These resuits suggested that triadimefon retarded the senescence of leaves in mung bean seedlings in terms of enhancing the protective ability of plant tissues against membrane lipid peroxidation.
Abstract (Browse 1896)  |  Full Text PDF       
The Chemical Constituents of Rabdosia coetsa
Author: LI Wen-Wu, LI Bo-Gang, DING Li-Sheng and CHEN Yao-Zu
Journal of Integrative Plant Biology 1998 40(5)
    From the leaves of Rabdosia coetsa (Buch.-Ham. ex D. Don) Ham, a new kauranoic acid, coetsanoic acid, along with eight known compounds, dihydrorabdoktmmin C, rabdoktmmin C, albopilosin A, maerocalyxin C, N-2'-hydroxy-eieosanoyl ( henicosanoyl-hexacosanoyl )-4-hydroxy- trans-8-sphingenine,ursolic acid, 2a, 3-dihydroxyursolie acid and daucosterol has been isolated. Its structure was established as 7a, 12a, 14-tri-hydroxy-15-oxo-ent-kauran-18-carboxy acid by spectroscopic means. The ceramides were isolated from the plants of Rabdosia for the first time.
Abstract (Browse 2155)  |  Full Text PDF       
Extracellular Calmodulin Stimulates the Transplasma Membrane Redox Reaction of Root Protoplasts in Zea mays
Author: SUN Yu, CHEN Jia and SUN Da-Ye
Journal of Integrative Plant Biology 1998 40(5)
    Using ferricyanide as the membrane impermeable electron acceptor, the effects of extracellular calmodulin on transplasma membrane redox reaction of the root protoplasts in Zea mays L. were studied. The calmodulin antagonists (calmidazolium, W7-agarose) and anti-calmodulin serum had inhibitory effect on the extracellular reduction of ferricyanide with their concentration that yielded 50% inhibition were 1.5 moL/L, 10 mol/L and 10 mg/L respectively. Inhibition of calmidazolimn could be restored by calmodulin completely. And the reduction of ferricyanide could be specifically stimulated by the exogerous purified calmodulin. These results suggested that transplasma membrane redox system of root protoplasts in Zea mays L. could be modulated by calmodulin outside the plasma membrane.
Abstract (Browse 1873)  |  Full Text PDF       
Studies of Signal Transduction of Salicylic Acid in Cucumber Cells
Author: LI Zhao-Liang, YUAN Yong-Bing, LIU Cheng-Lian and CAO Zong-Xun (Tsao T H)
Journal of Integrative Plant Biology 1998 40(5)
    For the first time, the signal transduction pathway of salicylic acid (SA) was investigated by using 3H-labelling, thin-layer chromatography and anion exchange column chromatography. It was found that SA stimulated the activity of membrane bound phospholipase C (PLC), accelerated the bm&down of phosphatidylinositol-4-monophosphate (PIP) and phosphatidylinositol-4,5-bisphos- phate (PIP2) and increased the levels of inositol-1,4-bisphosphate (IP2), inositol-1, 4,5-trisphos- phate (IP3) and diacylglycerol (DAG). These indicated that signal transduction of SA was probably accomplished through the mediation of phosphatidylinositide signal transduction system in cucumber ( Cucumis sativa L. ).
Abstract (Browse 1866)  |  Full Text PDF       
Studies on Root Regeneration from Antherderived Spear of Asparagus officinalis
Author: CHEN Sui-Yun, CHEN Yong-Zhe and REN Qun
Journal of Integrative Plant Biology 1998 40(5)
    There were two different stages during the root regeneration from the anther derived spear of Asparagus officinalis L. c v. Marry Washington 500, the root primordia differentiation, and the development stage of root primordia into young root. Differentiation of root primordia was enhanced by amplified nutrient supplement and lowered NAA concentration, both of which were favorable to callus formation and sustained decrease of endogenous ABA at the transection of the base of the spear. Moreover, further development of the root primordia was promoted by appropriate limitation of water supply and by effective aeration.
Abstract (Browse 1933)  |  Full Text PDF       
Optimization of Electrofusion Parameters and Interspecific Somatic Hybrid Regeneration in Citrus
Author: GUO Wen-Wu, DENG Xiu-Xin and SHI Yong-Zhong
Journal of Integrative Plant Biology 1998 40(5)
    This study was primarily attempted to optimize the electrofusion parameters using protoplasts isolated from cell suspension cultures of "Page" tangelo ( Citrus reticulata Blanco x C. paradisi Macf) and mesophyll protoplasts of rough lemon ( Citrus jambhiri Lush) as fusion partners. It was shown that the binuclear heterokaryons frequency reached 15% with the following parameters: alternate current (AC) 125 V/cm, AC time 60 s, direct current (DC) 1 250 V/cm, DC pulse width 50 s, DC pulse interval O. 5 s, No. of DC pulse 3. Considering the fact that different types of protoplasts have different specific weights, higher frequency of the binuclear heterokaryons was obtained by controlling the centrifugation time after fusion. The fusion products regenerated into plantlets after 3 to 4 months of culture. Chromosome counting of the root tips and morphological observation of the regenerants verified that 78% were tetraploids and the rest were diploids with the leaf morphology of mesophyll parent. Peroxidase (POX) isozyme and RAPD analysis indicated that interspecific somatic hybrids were obtained and an autotetraploid plant of mesophyll parent type was also verified.
Abstract (Browse 1912)  |  Full Text PDF       
Restriction Endonuclease Digestion of Amplification Products Generated by RAPD Technique in a Population of Glycine soja
Author: WEI Wei,ZHONG Min, WANG Hong-Xin, YUN Rui, HU Zhi-Ang and QIAN Ying-Qian
Journal of Integrative Plant Biology 1998 40(5)
    n a population of Glycine soja L., the polymorphic loci could be hardly detected by RAPD markers, using several primers. These non-polymorphic amplification products were cleaved by some restriction endonuclease, such as Msp , Hinf , Taq , EcoR , Sal , Dra and Hae . After cleaving, the digested amplification products were detected on polyacrylamide gel electrophoresis with silver staining. It was found that: 1 ) some restriction endonucleases could not, and some others could effectively digest the random amplication products of the DNAs of G. soja; 2) some endonucleases could produce polymorphic DNA fragments after digestion of the non-poly-morphic products, but others could not even after digestion; 3) non-polymorphic amplification products amplified by some primers could produce polymorphic DNA fragments after digestion, while those by other primers could not. It could be concluded that the restriction endonuclease digestion of amplification products could increase significantly detectability of polymorphic DNA by RAPDs technique.
Abstract (Browse 1926)  |  Full Text PDF       
Isolation of Soybean Kunitz Trypsin Inhibitor Gene and Its Application in Plant Insect-resistant Genetic Engineering
Author: GAO Yue-Feng, ZHU Zhen, XIAO Gui-Fang, ZHU Yu, WU Qian and LI Xiang-Hui
Journal of Integrative Plant Biology 1998 40(5)
    Soybean Kunitz trypsin inhibitor belongs to protease inhibitor family with insecticidal properties. The soybean Kunitz trypsin inhibitor gene was obtained from the template cDNA which was synthesized from the total RNA of the isolated soybean ( Glycine max L. ) immature cotyledons by PCR, and cloned into Sma I site of pBluescript KS( + ). DNA sequence analysis indicated that the cloned 663 bp DNA fragment carried the entire coding sequence, which encoded a precursor composed of 217 amino acids, including a signal peptide composed of 25 amino acids at the N terminus, the mature protein composed of 181 amino acids and an extra 11 amino acids at the C tenninus. A series of plant expression vectors have been constructed for tobacco (Nicotiana tabacum L. ), rice ( Oryza sativa L. ) and cotton ( Gossypium hirsutum L. ) transformation. The transgenic tobacco was confirmed by PCR and Southern blotting. Data from bioassays of insects showed that the transgenic tobacco displayed notably resistance to the larvae of Heliothis armigera Hubner compared with wild tobacco.
Abstract (Browse 2078)  |  Full Text PDF       
Studies on the Lignans of Polygonum orientale
Author: ZHENG Shang-Zhen, WANG Ding-Yong, MENG Jun-Cai and SEN XU Wei
Journal of Integrative Plant Biology 1998 40(5)
    Three lignans were isolated from the aerial parts of Polygonum orientale L. Their structures were elucidated as arctiin (1), lappaol B (2), orientalin (3) by means of the spectral data and chemical methods. Among them, 3 is a novel compound, compounds 1 and 2 were first found in this plant.
Abstract (Browse 2613)  |  Full Text PDF       
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