September 1996, Volume 38 Issue 9


          Research Articles
The Chemical Constituents of Anemone rivulari
Author: Peng Shu-lin, Ding Li-sheng, Wang Ming-kui and Chen Pei-qing
Journal of Integrative Plant Biology 1996 38(9)
    A new triterpenoid saponin, anemoside A. was isolated from Anemone rivularis Buch.-Ham. ex DC. Its structure was elucidated as 3-O-[-D-ribopyranosyl (13)--L- rhamnopyranosyl (1 - 2) --L-arabinopyranosyl]-oleanolic acid-E8-O--L-rhamnopyranosyl (1 4)--D-glucopyranoside by means of chemical methods and 1H-NMR. 13C-NMR, DEPT, MS spectral analyses.
Abstract (Browse 1825)  |  Full Text PDF       
Cloning and Structure Analysis of rbcS Gene from Wild Soybean
Author: Cao Kai-ming, Yuan Wei-ming, Zhan Shu-xuan, Gu Qi-min and Xu Bao
Journal of Integrative Plant Biology 1996 38(9)
    About 1 kb fragment of rbcS (ribulose 1, 5-bisphosphate carboxylase small subunit) gene in wild soybean (Glycine soja, Ji 50017) was amplified from total DNA by PCR assay. Sequence analysis of the fragment indicated that 1089 bp sequenced included the whole coding region for Rubisco small snbunit. The rbcS gene in wild soybean encoded a precursor composed of a transit peptide of 55 amino acids and a mature protein of 123 amino acids. There were two introns found in the rbcS gene as other dicotyledonous species previously sequenced. Comparison of DNA sequences showed high degree homology of rbcS genes between wild soybean and cultivated soybean (Glycine max var. wayne). Some changes of amino acids emerged from the diverse nucleotides did not affect the function of the small subunit. These results may contribute some basic data in molecular biology to study the origin and evolution of soybean.
Abstract (Browse 1989)  |  Full Text PDF       
RAPD Determinations on the Genetic Differentiation of Chinese Common Wild Rice
Author: Wang Zhen-shan, Chen Hong, Zhu Li-huang and Wang Xiang-kun
Journal of Integrative Plant Biology 1996 38(9)
    Ninety accessions which included Chinese common wild rice (Oryza rufipogon) from 8 provinces and traditional cultivars from lower and middle basins of Yangtze River, southeast of China and Yunnan Province as well as some commercial varieties were analyzed by RAPD with 24 primers. A scattered figure suggesting the indica-japonica and wild-domestication differentiations among 90 rice accessions was generated based on RAPD data. The results indicated that Chinese common wild rice, indica and japonica accessions were divided into 3 groups respectively. Chinese common wild rice were somewhat closer to the japonica type than the indica type.
Abstract (Browse 1760)  |  Full Text PDF       
A Palynological Survey of Neogene Strata in Xiaolongtan Basin, Yunnan Province of South China
Author: Wang Wei-ming
Journal of Integrative Plant Biology 1996 38(9)
    A palynological investigation of 24 samples from the representative Neogene strata of the Xiaolongtan Basin (2348N, 103llE) were novelly carried out. Pollen samples were collected from the strata of N1 (clay, sandstone and conglomerate), N2 (clay intercalated with thin bedded lignite at the lower part, and coal bed at the upper part), N3 (marl), and N4 (clay intercalated with thin bedded lignite) in ascending order respectively. Pollen flora was characterized by the predominant appearance of angiospermous elements, representing the parent plants of evergreen broad-leafed types. Variations in the pollen flora were recognized between sample 15 and 16, as more herbaceous pollen of the Compositae types increasing upward in the strata, while certain woody types were minimized or even disappeared, thus dividing the pollen flora into two subzones. According to the current study, the geological age of the flora was suggested to be Middle-Late Miocene (Subzone A) to Early Pliocene (Subzone B), and the inferred vegetational type was evergreen broad-leafed forest, accompanied by a few deciduous elements.
Abstract (Browse 1865)  |  Full Text PDF       
Calcium Effect on the Process of Cold-Hardening of Rice Seedlings
Author: Li Mei-ru, Liu Hong-xian, Wang Yi-rou and Zeng Shao-xi
Journal of Integrative Plant Biology 1996 38(9)
    Rice seedlings for cold-hardening were germinated from seeds treated with or without immersion in 30 mmol/L of CaCl2 solution. Change of membrane system protectivity in leaves at various periods (viz: following cold-hardening, chilling stress and on the 3rd day of recovery); the ratio of seedling survival and the ability of recovery were investigated. The results showed that cold-hardening increased the contents of endogenous antioxidants (the reduced form of glutathione, GSH; ascorhic acid, ASA), SOD activity and the content of heat stable protein in soluble protein. Cold-hardening combined with the CaC12 treatment of seeds enhanced the above-mentioned effects of cold-hardening, and obviously increased the activities of catalase (CAT) and peroxidase (POD). Both cold-hardening with or without the CaCl2 treatment reduced the declining degree of GSH, AsA content, the SOD activity and the heat stable protein content caused by chilling stress, augmented the increase in the level of GSH, AsA, CAT, SOD, POD and heat stable protein in the recovery periods. The CaC12 pretreatment especially had more effect on the level of heat stable protein, augmenting the A protein to rise to the level in normal seedlings. Furthermore, cold-hardened seedlings from seeds pretreated with CaClz grew faster and better in the recovery period of chilling stress than those from seeds without CaClz pretreatment. This promotive effect of CaC12 treatment on seeds prior to cold-hardening of seedling could be associated with an aug- mentation of membrane protectivity induced by the combined treatment.
Abstract (Browse 1814)  |  Full Text PDF       
Plant Regeneration from Barly (Hordeum vulgare) Immature Inflorescences Cryopeserved by Vitrification
Author: Wang Jun-hui, Yan Qing-feng and Huang Chun-nong
Journal of Integrative Plant Biology 1996 38(9)
    Vitrification is a new technique of cryopreservation. Immature inflorescences of barley (Hordeum Vulgate L. ) were precultured in medium supplemented with 0. 5 mol/L sucrose for 2 days and dehydrated in PVS2 at 0 for 5 min before plunged into liquid nitrogen. After rapid warming and unloading, the survival of three cultivars "81Gl", "Gebeina", and "Xu 9" was 100%, 82.5% and 50% respectively. Callus formation and plant regeneration was achieved from recovered inflorescences. The non-precultured samples could not survival in liquld nitrogen. Desiccation-tolerance and freeze-tolerance affected plant survival. Vitrification shared the advantages of high levels of survival, short period of lag stage and complete viability of cryopreserved organs.
Abstract (Browse 1841)  |  Full Text PDF       
Relation Between Desiccation Sensitivity and Membrane Permeability of Developing Clausena lansium Seeds
Author: Jiang Xiao-cheng, Fu Jia-rui and Huang Sheng-qin
Journal of Integrative Plant Biology 1996 38(9)
    Desiccation sensitivity and its relation to membrane permeability of the embryonic axes of the developing wampee (Clausena lansium (Lour.) Skeels) seeds were studied by measuring the changes in electrolyte leakage, germination and vigor index after the embryonic axes were rapidly air-dried to various water contents. During development, the fresh and dry weight per seed reached nearly maximum value at 72 d after anthesis, but the dry weight per embryonic axis continuously increased until 85 d after anthesis. The embryonic axes acquired the full capacity for germination at 58 d after anthesis and their vigor index continuously rose up from 51 to 92 d after anthesis. The electrolyte leakage of the developing the embryonic axes linearly declined to the minimum value at 72 d after anthesis and then went up again. The electrolyte leakage of the embryonic axes was higher than that of the whole seeds at the same time. The immature embryonic axes did not germinate completely, while mild desiccation could improve their viability. Any degree of desiccation decreased the vigor index of the embryonic axes which have reached physiological maturation and the decline of vigor index was corresponded to the increase of electrolyte leakage. According to this experiment, the authors concluded that wampee seeds did not gain desiccation-tolerance which was a characteristic of orthodox seeds during development. High water content was essential for maintaining membrane integrity and stabiligy of matured wampee seeds. The injury of seed viability during dehydration could be estimated by using the electro-conductivity method.
Abstract (Browse 1806)  |  Full Text PDF       
Nucleotide Sequence of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Large Subunit Gene from Millet (Setaria italica)
Author: Zhao Yin-suo, Qiao Xiao-yan, Wu Nai-hu and Wu Xiang-yu
Journal of Integrative Plant Biology 1996 38(9)
    The 3.0 kb Bgl + Xba restriction fragment of millet (Setaria italica) chloroplast genome containing rbcL gene had been cloned into pBluescript SK (һ) vector, then the restriction map and the 1990 bp complete nucleotide sequence was determined. The 1431 bp coding region of the gene consists of 476 amino acid residues with a predicted molecular weight of 52679 D. The 389 bp 5 upstream region has the putative -10 box, -35 box and SD sequence, similar to that of procaryotes. The 170 bp 3 downstream region contains three stem-loop structures. Comparison of the rbcL gene sequences between C4 plants and several C3 plants reveals no difference in the coding region, promoter and 3' downstream region. It might be concluded that the rbcL gene sequence has no relation with its cell-specific expression.
Abstract (Browse 1714)  |  Full Text PDF       
Transgenic Potato with PVY Coat Protein Gene and Its Small-Scale Field Test
Author: Song Yan-ru, Ma Qing-hu, Hou Lin-lin, Zhang Li-zhi, Yang Wen-yu, Peng Xue-xian and Wang Hai-yun
Journal of Integrative Plant Biology 1996 38(9)
    Potato virus Y (PVY) infection may cause a severe yield depression up to 80%. To develop the potato (Solanum tuberosum L. ) cultivars that resist PVY infection is very crucial in potato production. The authors have been cloned the coat protein gene of PVY from its Chinese isolate. A chimaeric gene containing the cauliflower mosaic virus 35S promoter and PVY coat protein coding region was introduced into the potato cultivars Favorita, Tiger head and K4 via Agrobacterium tumefaciens. Results from PCR and Southern blot analysis confirmed that the foreign gene has integrated into the potato chromosomes. These transgenic potato plants were mechanically inoculated with PVY virus (20 mg/L). The presence of the virus in the potato plants was determined by ELISA and method of back inoculation into tobacco. The authors observed a drastic reduction in the accumulation of virus in some transgenic potato lines. Furthermore, some transgenic potato lines produced more tubers per plant than the untransformed potato did, and the average weight of these transgenic plant tubers was also increased. In the field test, the morphology and development of these transgenic potato plants were normal, 3 transgenic lines of Favorita exhibited a higher yield than the untrasformed virus-free potato with an increase ranged from 20% to 30%. From these transgenic lines, it will be very hopeful to develop a potato cultivar which not only has a significant resistance to PVY infection, but also a good harvest in potato production.
Abstract (Browse 1826)  |  Full Text PDF       
Primary Analysis of Components in Different Soybean Nodulating Mutants and their Effects on Nitrate Reductase Activity and Nodulation
Author: Li Zhi-zheng and Gong Song-fu
Journal of Integrative Plant Biology 1996 38(9)
    The water extracts of leaves and roots from supernodulating soybean (Glycine max (L.) Merr. ) nts 382 and nonnodulating soybean Nod 49 have been chromatographed using filtering method through the column (25 cm 2 cm) Sephadex G25 and 4 fractions, namly, nts 382 (Nod 49) F1, nts 382 (Nod 49) F2, nts 382 (Nod 49) F3, and nts 382 (Nod 49) F4 could be distinguished according to nitrate reductase (NR) activities inhibited by the eluate. The inhibition of NR activity by the noninoculated nts 382 F2 and the nts 382 F4 in vitro were much stronger than that by the inoculated nts 382 F2 and nts 382 F4. On the contrary, the obvious inhibition of NR activity in vitro by the noninoculated Nod 49 F2 and Nod 49 F4 were substantialy strengthed again by the innoculated Nod 49 F2 and Nod 49 F4. The facts indicated that the quantity of NR inhibitors in the leaf cells of soybean nts 382 reduced after the inoculation but was that in the inoculated Nod 49 leaf cells further more accumulated. Both nodulations assays, the nodulation of soybean "Bragg " injected with inoculated nts 382 Fl, nts 382 F2, nts 382 F3 and nts 382 F4 from leaves and roots and the nodulation of soybean nts 382 injected with inoculated Nod 49 F2, Nod 49 F3 and Nod 49 F4 from leaves only showed that nts 382 Fl and nts 382 F2 increased nodules of soybean "Bragg" by 1 to 3 times but nts 382 F3 and nts 382 F4 did not. Inhibition of soybeannts 382 nodulation by inoculated Nod 49 F2 Nod 49 F3 and Nod 49 F4 expressed that the Nod 49 F4 only inhibited the nodulation strongly by one time in the experiments with nts 382 plants with leaves, and by 15 times in the experiments with nts 382 plants without leaves at 10 d of inoculation and injection and this inhibition was nonreversible even after stopping injection from the 11th day to the 15th day after inoculation.
Abstract (Browse 1758)  |  Full Text PDF       
The Effect of Urea on the Activity and Stability of Metallocluster of the MoFe Protein of Nitrogenase from Azotobacter vinelandii
Author: Wang Zhi-ping, Zhang Hui-miao, Huang Ju-fu, Luo Ai-ling and Li Jia-ge
Journal of Integrative Plant Biology 1996 38(9)
    When the MoFe protein from Azotobacter vinelandii was treated with more than 0.5 mol/L of urea anaerobically, the C2H2-reduction activity of the treated protein was exponentially decreased. However, the activity could be significantly restored after the treated protein was diluted with the buffer system and followed by incubation at 15 . The urea had remarkable enhancement on chelation of Fe atoms from the reducted MoFe protein and the P-cluster-deficient MoFe protein by O-phenanthroline (O-phen), respectively. The migration of the reducted MoFe protein on the urea-gradient gel electrophoresis did not significantly change from 0 to 1.5 mol/L urea , linearly became smaller from 1.5 to 5.0 mol/L urea, and reached a stable state from 5.0 to 8.0 mol/L urea. The results indicated that: (1) The effect of urea on the activity and the stability of metallocluster of the MoFe protoin was mainly attributed to the conformational change of the protein in urea, moveover, the effect of urea on the metallocluster might not be all the same if the states of MoFe protein were dif ferent; (2) There was a close relationship between the conformation and the metalloclusters of MoFe protein; (3) In the course of the denaturation, the decrease in the activity of MoFe protein probably happened prior to the conformational change of the whole molecule.
Abstract (Browse 1932)  |  Full Text PDF       
Confocal Microscopic Observations on Microtubular Cytoskeleton Changes During Megasporogenesis and Megagametogenesis in Phaius tankervilliae (Aiton) Bl.
Author: Ye Xiu-lin, Yeung Edward, Xu Shi-xiong (S. Y. Zee) and Tong Sui-hai (Tung Shiu-Hoi)
Journal of Integrative Plant Biology 1996 38(9)
    In nun orchid (Phaius tankervilliae (Alton) B1. ) embryo sac development follows the monosporic pattern. Changes in the pattern of organization of the microtubular cytoskeleton during megasporogenesis and megagametogenesis in this orchid were studied using the immunofluorescence technique and eonfocal microscopy. At the initial stage of development the microtubules in the arehesporium were randomly oriented into a network. Later the archesporial cell elongated to form the megasporocyte. The cytoskeleton in the elongated megasporoeyte was radially organized in which microtubules extending from the nuclear envelope to the peripheral region of the cell. The megasporoeyte then underwent meiosis 1 to form a dyad. The dyad cell at the chalazal end was larger than the cell at the micropylar end. Microtubules in the dyad cell were radially oriented. The dyad underwent meiosis to give rise to a linear array of four megaspores (i. e. tetrad formation). The chalazal-far most megaspore survived and became the functional megaspore, which contained a set of randomly oriented microtubules. The microtubules in the other 3 megaspore disappeared as the cells degenerated. The functional megaspore then underwent mitotic division giveing rise to a 2 nucleate embryo sac. The nuclei of the 2-nucleate embryo sac were separated by a set of longitudinally oriented microtubules which ran parallel to the long axis of the embryo sac. Each nucleus in the embryo sac was surrounded by a set of perinuelear microtubules. The gnucleate embryo sac again underwent mitotic division to form a 4-nucleate embryo sac. The division of the two nuclei was synchronous. But the orientation of the division plan of the two spindles was different (i. e. the spindle microtubules at the chalazal end ran parallel with the long axis of the embryo sac and those at the mieropylar end ran at right angle to the axis of the embryo sac). The 4 nuclei of the 4-nucleate embryo sac were all tightly surrounded by randomly oriented microtubules. Later the paired nuclei at the micropylr end and at the chalazal end as well underwent mitotic division in seguence. At this time when the embryo sac had reached the 8-nucleate embryo sac stage. The pattern of organization of the microtubules was very complex. Initially the nuclei were surrounded by a set of randomly oriented microtubules, but after the two polar nuclei had moved to the central region of the embryo sac, three different organizational zones of microtubules appeared, viz: a randomly oriented set of microtubules surrounding each nucleus in the chalazal zone: a set (in the form of a basket) of cortical microtubules which surrounded the vacuoles and the two polar nuclei in the central zone and a loosely knitted network of microtubules surrounding the nucleus that later became the egg cell nucleus in the micropylar zone. The two nuclei that would become the nuclei of the synergids were surrounded by a set of more densely packed mierotubules. Towards far the most micropylar end some microtubules formed thick bundles. The site of appearance of these thick bundles coincided with the site of development of the filiform apparatus. The pattern of microtubule organization after cellularization (i. e. at the beginning of embryo sac maturation) did not change much. The author's results indicated that various patterns of microtubule organization observed in the developing embryo sac of nun orchid reflected the complexity and dynamism of the embryo sac.
Abstract (Browse 1995)  |  Full Text PDF       
Direct Measurement of the K+-Channel and Its Ion Selectivity in the Thylakoid Membrane from Spinach (Spinacea oleracea L.) Leaves
Author: Li Le-gong, Yan Ji-qiong and Tang Zhang-cheng
Journal of Integrative Plant Biology 1996 38(9)
    Thylakoid vesicles were purified from spinach (Spinacea oleracea L. ) leaves by sucrose density gradient centrifugation and incorporated into planar lipid bilayers by stirring. At least one type of voltage-dependent K+ single-channel currents was found. Its conductance (between +60 mV and C60 mV ) was about 55 pS in symmetrical (cis: trans) 250 mmol/L KC1. This channel was sensitive to TEA (tetraethylammonium chloride) and the permeability ratio (PK+/PCl-) was about 14. 9. The selectivity of 55 pS channel determined from both reversal potentials under bi-ionic conditions or from conductance measurements in symmetrical solutions, was in the seguence of K+Na+Li+ NH4+ Cs+. This potassium channel could act as involved in charge-balancing during light-driven proton uptake by thylakoid.
Abstract (Browse 1718)  |  Full Text PDF       
Effects of Nifedipine on Pollen Germination, Pollen Tube Growth and Division of Generative Nucleus in Nicotiana tabacum
Author: Fan Liu-min, Yang Hong-yuan and Zhou Chang
Journal of Integrative Plant Biology 1996 38(9)
    The deals with the effects of nifedipine (Nif), a Ca2+ channel blocker of rather high specificity, on pollen germination, pollen tube growth and division of generative nucleus (GN) in experimentlly germinated pollen tubes of Nicotiana tabacurn L. Pollen germination was inhibited by the addition of 10-4 mol/L Nif whereas no significant inhibition by 10-710-5 mol/L Nif was observed. The effects of Nif on pollen tube growth were related to its concentration and duration of treatment. At the earlier stage, tube growth was promoted at the lower concentrations (10-710-5 mol/L), but was significantly inhibited at a concentration of 10 4 mol/L Nif. With increasing time of culture, even the lower concentrations also became harmful; the stronger the concentration, the earlier the transition from promotion to inhibition. Generally, inhibition of tube growth occurred within 24 hours of culture with different extent in various concentrations. Moreover, higher concentrations also tended to disturb tube morphology and cytoplasmic streaming. Nif was observed to perturb GN division at various concentrations, either blocked it completely at 10-4 mol/L, or only delayed it at 10-710-5 mol/L. The dynamics of membrane-associated calcium in pollen tubes was tested with chlorotetracycline (CTC). With increasing time of culture and escalating Nif concentration, CTC fluorescence weakened gradually, indicating that the physiological effects of Nif is mediated by its in hibition on Ca2+ channel activities.
Abstract (Browse 1938)  |  Full Text PDF       


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