June 1997, Volume 39 Issue 6


          Research Articles
Distribution of Repetitive DNA Sequences Derived from Rice Genome in Genus Oryza and Family Poaceae
Author: Han Ai-dong, Lin Jian-dong, Miao Ru-huai, Yu Hao and Liu Liang-shi
Journal of Integrative Plant Biology 1997 39(6)
    Some repetitive sequences have been cloned from rice genome by DNA renaturation kinetics. Using them as probes, Southern blotting analyses in 30 species of Oryza and Poaceae were carried out. The authors' results indicated that hybridization band patterns correlated with relationship of plants. The use of these repetitive sequences as probes in phytogenetic classification is considered. The fragment of pRRD9 also showed polymorphisms among some ecological types of wild rice ( O. rufipogon), suggesting that it could be used as a probe in the study of rice evolution.
Abstract (Browse 1697)  |  Full Text PDF       
Human Defensin-1 (HNP-1) Expression in Tobacco: A Preliminary Observation on Anti-TMV Activity of the Transgenic Tobacco Plants
Author: Zhang Man-chao and Zheng Guo-chang
Journal of Integrative Plant Biology 1997 39(6)
    Human defensin-1 (human neutrophil peptide-1, HNP-1) is one of the small peptides found recently in human neutrophils with high antimicrobial activity of broad spectra. In this work, HNP-1 plant expression vectors with different promoter, with prepropiece and without the pre- and most part of propiece of HNP-1 cDNA have been constructed, and a serial of agrobacteria have been reconstructed using these vectors by tri-gameting. Tobacco ( Nicotiana tabacum L. ) leaf discs were infected with these reconstructed agrobacteria and plants regenerated after about 1 month. Southern blotting analysis showed that HNP-1 cDNA had been integrated in tobacco genome, but ELISA analysis indicated that only transgenic plants infected with agrobacteria contained HNP-1 expression vectors with an improved CaMV 35S promoter and without pre- and most part of the propiece of HNP-1 cDNA can express HNP-1 at Iow level. Information from electron microscopic observation and external feature of the plants showed that these plants had the ability to defend plants from TMV infection at a certain concentration (0.2 to 0.8 mg/L). It was concluded that it was a prerequisite to introduce an efficient promoter to an expression vector for an effective expression of a foreign cDNA, and it was necessary to delete the pre- and most part of the propiece of an animal cDNA for its expression in plants. Furthermore, HNP-1 cDNA transferring into plants could create a novel method to produce antipathogenic crops.
Abstract (Browse 1655)  |  Full Text PDF       
Relationship Between Ultrastructural Changes and Zearalenone in Triticum aestivum During Vernalization
Author: Zhao De-gang and Meng Fan-jing
Journal of Integrative Plant Biology 1997 39(6)
    The zearalenone (ZEN) levels and cell uhrastructure in the shoot apex of winter wheat ( Triticum aestivum L. cv. Yanda 1817) during vernalization were studied by immunosorbent assays and electron microscopy. The main results showed that the ZEN content in seed embryo increased during vernalization, and reached the maximum at week 4, and decreased thereafter. The obvious uhrastmctural changes of apex appeared after 4 weeks of low temperature. The apex cells enriched organelles, such as plastids, mitochondria, rough endoplasmic reticula. A lot of plastids and mito- chondria were irregular being budding or dumbbell shaped. The invagination of the plasma mem- brane and the nuclear substance transpassing though cell wall were visualized. Application of malathion (MAL), an inhibitor of ZEN biosyntheses, inhibited the formation of ZEN during vernalization; also affected the ultrastructure of apex cells of the induced seedling. The application of exogenous ZEN might promote the development of plastid, mitochondrion, rough endoplasmic reticulum and dictyosome.
Abstract (Browse 1680)  |  Full Text PDF       
Recent Advances in Molecular Biological Studies on Disease-Resistant Genes in Plants
Author: Zhu Guo-feng, Qu Li-jia, Gu Hong-ya and Chen Zhang-liang
Journal of Integrative Plant Biology 1997 39(6)
    As the rapid development of molecular biological techniques, scientists have achieved a great deal in probing the plant-pathogen (bacteria and fungi) interaction at molecular level. Nine plant disease-resistant genes (R genes) have been cloned and they can be divided into four classes. The most important characteristic of R genes is that they encode a leucine rich repeat motif or a serine-threonine protein kinase motif. They might involve in the specific recognition between host and pathogen, activating the subsequent signal transduction which leads to the hypersensitive response in the host plant. The common feature of coding for leucine rich repeat motif and tandem arraying in the chromosomes seem indicating the common ancestral for these genes. This paper also summarizes the current work on studies of anti-fungal and- bacterial genes at molecular level in the authors* laboratories.
Abstract (Browse 1932)  |  Full Text PDF       
Establishment of Transfer Functions of the Pollen-Climatic Factors in Northern China and the Quantitative Climatic Reconstruction at DJ Core
Author: Song Chang-qing and Sun Xiang-jun
Journal of Integrative Plant Biology 1997 39(6)
    The 2.4 meter-long core was extracted from the Diaojiao lake (41º18∩N, 112º21∩E) at the foot of the northern part of Daqingshan Mts. Pollen analysis from collections subsampled in the laboratory at 2 cm intervals, revealed plentiful pollen and spores from over 10 arboreal genera, including Pinus, Betula, Picea, Abies, Carpinus, Quercus, Ulmus and more than 20 non-arboreal genera, mainly of Artemisia, Labiatae, Nitraria, Polygonaceae, Ranunculus, Thalictrum, Umbel- liferae, Caryophyllaceae and Cyperaceae. Fern spores, aquatic pollen and algae were also observed in some parts of the core. The transfer functions were established by the stepwise regression analysis using the climatic factors and 13 pollen taxa. The different Fl and F2 value were used as the thresh- old value of F test (i. e. used for selecting and deleting factors). Each regressed equation was obtained from 70 times of calculations with a step-wise increase of 0.1 for Fl and F2 and those having the smallest regression deviation and the largest multiple correlation coefficient were the final four transfer functions. Substituting the pre-factor obtained from the stratigraphic sampled into the regression equations, the estimates of temperature and precipitation in January and in July, and annual mean temperature values could be calculated. Some climatic stages were inferred from total pollen influx and pollen percentage from the core using a transfer function: humid-cool (from 10 000 to 7 800 a BP), arid-cold (9 200 to 7 900 a BP), arid-warm (7 900 to 7 100 a BP), humid-warm (7 100 to 4 400 a BP), arid-warm (4 400 to 3 000 a BP), arid-cold (3 000 to 2 100 a BP). The highest annual mean temperature during Holocene was ca. 4 ⊥ higher and the lowest was ca. 2 ⊥ lower than the present temperature. Annual precipitation was 250 mm higher and 300 mm lower than the present.
Abstract (Browse 1763)  |  Full Text PDF       
Changes in Stomatal Conductance, Transpiration and Water Use Efficiency of Ten Species Experienced in High CO2 Concentrations in Biosphere 2
Author: Jiang Gao-ming, Lin Guang-hui and Bruno D V Marino
Journal of Integrative Plant Biology 1997 39(6)
    The stomotal conductance, transpiration and water use efficiency (WUE) were measured using a LI-6400 portable photosynthesis system for 5 tropical rain forest species and 5 desert species in Biosphere 2, USA. All the species have experienced in very high CO2 ( > 2 200 米mol• mol- 1 ) for more than 4.5 years. The results showed that the stomatal conductance and transpiration of rain forest species decreased from ( 127.4 ㊣ 65.6) and (2.04 ㊣ 0.61 ) mmol• m- 2•s- 1 to (61.3 + 30.5) and ( 1.54 ㊣ 0.65 ) mmol• m-2• s -1 respectively, while WUE increased from (2.90 ㊣ 0.55) to (8.45 ㊣ 2.71) 米mol CO2 •mmo1-1 H2O, with CO2 increasing from 350 每 400 to 700 每 820 米mol• mol-l. For the desert species, stomatal conductance and transpiration decreased from respectively (142.8㊣94.6) and (2.09㊣0.71) mmol•m-2•s-1 to (57.7㊣35.8) and (1.36㊣0.52) mmolm-2•s-l, but WUE increased from (4.69 ㊣ 1.39) to (9.68 ㊣ 1.61) 米mol CO2•mmo1-1 H2O, with the CO2 increase from 320 - 400 to 820 每 850 米tmol• mol- 1. The stomatal conductance, transpiration and WUE were less influenced by light intensity under high CO2 than low CO2 concentrations. Most rain forest species reached their light saturation points at light intensity of 500 米mol• m-2•s-1, while desert species at 1 000 米mol•m-2•s-1. Among different species, the desert C3 tree, Nicotiana glauca Grah., had the highest decrease in stomatal conductance and transpiration and the highest increase in WUE, by 78%, 69% and 310% respectively. The enhancement of increasing CO2 to the stomatal, transpiration and WUE of species with different photosynthesis pathway and life forms in Biosphere 2 could be concluded as: C3 species > C4 species, and desert C3 species > rain forest C3 species.
Abstract (Browse 2167)  |  Full Text PDF       
Promotive Effect of Triadimefon on Drought Resistance of Rice Seedlings
Author: Guo Zhen-fei, Lu Shao-yun and Li Ming-qi
Journal of Integrative Plant Biology 1997 39(6)
    Triadimefon increased the relative water content and decreased malondialdehyde (MDA) content under osmotic stress of 每 0.5 MPa induced by PEG-6000. The activities of peroxidase (POD) and catalase (CAT) were induced by triadimefon at a concentration of 75 mg/L by root treatment under osmotic stress, but the superoxide dismutase (SOD) activity remained unchanged. Addition of cycloheximide, an inhibitor of protein synthesis, indicated that the activation of POD induced by triadimefon was due to the promotive effect of de novo enzyme synthesis. No effect of triadimefon on POD isozyme pattern was demonstrated on PAGE.
Abstract (Browse 1721)  |  Full Text PDF       
In Vitro Enzymological Studies on Anabolism of 2,4-diaminobutyric Acid in Lathyrus sylvestris
Author: Mu Xiao-min, Shen Li-ming and Wu Xian-rong
Journal of Integrative Plant Biology 1997 39(6)
    An in vitro synthetic reaction system was established with 2,3-3H-aspartic acid (Asp) as a substrate and the homogenate of fiatpea ( Lathyrus sylvestris L. ) leaves as the crude enzyme extract. The results showed that 3H-Asp was incorporated into 2,4-diaminobutyric acid (DABA). The incorporation was inhibited by the addition of glutamic acid (Glu). 3H-Asp was also incorporated into DABA after the cmde enzyme was dialyzed, indicating that Asp as a substrate for DABA synthesis was catalyzed by a group of enzymes which converted Asp to DABA in flatpea. From the in vitro reactions it was proved that DABA and 污-aminobutyric acid (GABA) could not be mutually substituted as substrates.
Abstract (Browse 1759)  |  Full Text PDF       
A Method for Purification of Antibodies with Protein Immobilized on Nitrocellulose Membrane
Author: Luo Ai-ling, Liang Zheng and Maurice S B Ku
Journal of Integrative Plant Biology 1997 39(6)
    A improved method for affinity purification of antibodies has been developed in which protein was immobilized on nitrocellulose membrane. So that antibodies could be specifically bound to its antigens rapidly, efficiently and precisely.
Abstract (Browse 1657)  |  Full Text PDF       
Relationship Between Endogenous Ethylene and the Development and Operation of the Alternative Respiratory Pathway in Aged Potato Tuber Slices
Author: Liang Wu-sheng, Wen Jiang-qi and Liang Hou-guo
Journal of Integrative Plant Biology 1997 39(6)
    The relationship between endogenous ethylene and the development and the operation of the alternative respiration pathway in aged potato (Solanum tuberosum L. ) tuber slices were investigated. During the 24 h aging period under 30 ⊥, along with the great increase of the total respiratory rate (Vt) of the slices, the alternative pathway capacity (Valt) developed continuously, and the Valt/Vt values kept increasing as well. Both the alternative pathway activity (老Valt) and its contribution to Vt(老Valt/Vt) also increased gradually before 12 h of aging, but kept constant from 12 h to 24 h. The time course of the endogenous ethylene production of the aged slices was similar to the changing trends of Valt and Valt/Vt, but different from those of 老Valt and 老Valt/Vt values. ACC and Cu2 + treatment which stimulated ethylene production of the aged slices enhanced their Valt and Valt/ Vt values, Co2 + and Ag+ treatment resulted in a decreased effect. However, all the above treat ments of ethylene-related agents could not alter the continuously decreasing trend of the 老 value of the alternative pathway of the aged slices. And their effects on 老Valt and 老Valt/Vt values were only observed before 12 h of aging. These results suggested that the endogenous ethylene was essential to the development of the capacity of alternative pathway, but could only slightly influence the operation of the activity of the alternative pathway in aged potato tuber slices.
Abstract (Browse 1717)  |  Full Text PDF       
Peroxidation Damage of Oxygen Free Radicals Induced by Cadmium to Plant
Author: Huang Yu Shan, Luo Guang-hua and Kwan K M Frances
Journal of Integrative Plant Biology 1997 39(6)
    Two cultivars of Festuca rubra L., namely metal tolerant "Merlin" and metal sensitive "S59", were treated with culture media containing Cd2 + at various concentrations ranging from 0 to 2.5 mmol/L. The biomass of the grass seedlings of both cuhivars was significantly dropped in connection to increase in metal concentration and treatmental time. The degree of inhibition by 0.5 mmol/L Cd2+ in "Merlin" was quite similar to that caused by 0.25 mmol/L Cd2+ in "S59" ,indicating that "Merlin" was relatively resistant to metal stress. Accompanying with the inhibition of growth by Cd2 + , malonyl dialdehyde (MDA) content and peroxidase (POD) activity measured in the seedlings increased drastically, while the activities of catalase (CAT) and superoxide dismutase (SOD) decreased with significant drop in total protein content. The remarkable enhancement of MDA in the grass seedlings appeared to be an obvious physiological response due to Cd2 + treatment. When plants treated with 2.5 mmol/L Cd2+ for 6 days,MDA increased by 109% and 143% in "Merlin" and "S59", respectively. There was a close positive correlation between MDA content and Cd2 + concentration, and a negative correlation between MDA and plant growth. As slight decrease in SOD activity was found accompanied by a drastic elevation of MDA content. This observation may imply that free radical and MDA may play an important role in the Cd2 + led toxicity in plants.
Abstract (Browse 1840)  |  Full Text PDF       
The Cytoultrastructure of Nostoc flagelliforme Under Cultured Condition
Author: Liu Ming-zhi and Pan Rui-chi
Journal of Integrative Plant Biology 1997 39(6)
    Cellular ultrastructure of Nostocflageliforme Born. et Flab. under different growth conditions have been observed in order to study the characteristics of growth and development and the mechanism of gelatinous sheath formation. All kinds of cells were in an active metabolic state of growth with regular cell division in distilled water after two hours. The formation of gelatinous sheath was associated with the cell wall of the vegetative cells and a special type of macrocells, both of which were physiologically active at a culture-medium pH of 7.0. In an acid pH environment, however, the vegetative cell wall became inactive, no macrocells appeared but instead, a large number of a special type of microcells were observed.
Abstract (Browse 1731)  |  Full Text PDF       
Ultrastructural Study on the Ontogeny of Minor Veins and Associated Sheath Tissues in the Maize Leaf Blades
Author: Ouyang Xue-zhi and Xie Shao-ping
Journal of Integrative Plant Biology 1997 39(6)
    The origin, early development and ultrastrueture of the minor vein procambium and associated bundle sheath cells were studied in the leaf blades of maize ( Zea mays L. ). Apparently three contiguous cells in the middle layer of ground meristem apparently might be involved in the initiation of a minor vein and bundle sheath. The minor veins were formed from two unequal periclinal divisions of the inner cell, which gave rise to three smaller cells, one centrally located procambial initial cell, two bundle sheath cell precursors adaxially and abaxially located in respect. One of the two lateral cells immediately adjacent to either side of the inner cell directly differentiated into a precursor also divided radially, producing two abaxial bundle sheath cells. At the early stage of minor vein development, the procambial cells were essentially similar in ultrastructure to their precursor procambial initial cell and had all the protoplasmic constituents typically encountered in the meristematic cells. They contained a large nucleus with aggregates of heterochromatin and a dense cytoplasm rich in ribosomes. Proplastids possessed a simple fine structure with a few or without any internal membranes. Short segments of rough endoplasmic reticulum were sparsely distributed in the cytoplasm or associated with the plasmodesmata and with the outer membrane of the nuclear envelope. In many cases, the cytoplasm contained a few small vacuoles, many of which were in close association with the dilated endoplasmic reticulum. Numerous plasmodesmata were also observed in the walls between procambial cells and between procambium or procambial initial and surrounding cells. At the later stage of procambium development, some procambial cells showed an increased thickness of the cell wall, which was accompanied by a substantial decrease in the number of plasmodesmata between these cells, while other procambial cells transformed into the highly vacuolated parenchymatous state. The bundle sheath cells began to differentiate very shortly after vein initiation. The differentiation of the meristematic sheath cells into the highly vacuolated, parenchymatous photosynthetic cells was a gradual process. That was accompanied by the transition of proplastid to chloroplast, cell volume enlargement, and formation of large central vacuoles. The latter was also associated with fusion of small vacuoles as well as invagination of cytoplasmic material, including small vesicles, mimicking phagocytosis. The results indicated that adaxial and abaxial bundle sheath cells of the minor veins were more closely related ontogenetically to the initial procambial cell than were the lateral bundle sheath cells which were closely related ontogenetically to the mesophyll cells in the middle layer of the leaf.
Abstract (Browse 1943)  |  Full Text PDF       
Photoreduction of Cytochrome b559 in the Photosystem j Reaction Center Complex
Author: Lu Rong-he, Tang Chong-qin, Kuang Ting-yun and Tang Pei-song
Journal of Integrative Plant Biology 1997 39(6)
    The isolated and purified photosystem j (PS j ) reaction center D1/D2/Cyt b559 complex was taken as the experimental system. It was observed that under anaerobic conditions, cytochrome b559 (Cyt b559) could be reduced by exposure to strong illumination, suggesting Cyt b559 could accept electrons directly from reduced pheophytin (Pheo-). And the photoreduction of Cyt b559 was irreversible. When the isolated D1/D2/Cyt b559 complex reconstituted with exogenous secondary electron acceptor 2,6-dimethyl-benzoquinone (DMBQ), the photoreduction of Cyt b559 was delayed in the function of illumination time. Meanwhile, the electrons transferred mainly through DMBQ and photoreduced Cyt b559 could be partially reoxidized in the dark incubation following illumination. It was concluded that the quinone-independent electron transfer via Cyt b559 was a new, secondary electron pathway, which represented one of the protective pathes for PS j reaction center to dissipate excess excitation energy.
Abstract (Browse 1734)  |  Full Text PDF       
The Specific Proteins of Sperm Cells of Lilium davidii
Author: Tang Jian, Zhu Guang-lian and Cao Zong-xun (Tsao T H)
Journal of Integrative Plant Biology 1997 39(6)
    Viable generative cells and sperm cells of Lilium davidii Duch. were successfully isolated and purified with osmotic shock method and Pereoll density gradient centrifugation. Total proteins were extracted from sperm cells, generative cells and leaves of L. davidii. Protein patterns were compared on two-dimensional-PAGE. There were apparently more protein dots on the gel from sperm cells than on that from generative cells, especially in the alkaline region. Viable leaf protoplasts were isolated with multi-enzyme mixture, followed by centrifugation. The plasma membrane proteins of sperm cells, generative cells and intact leaf protoplasts were compared, using NHS-biotin (N-hydroxysuccinimido-biotin) labeling method. Horseradish peroxidase labeled streptavidin and the sub-strate 4-Chloro-l-naphthol were used to detect membrane proteins during Western blotting. Two proteins with molecular weights of 46 kD and 50 kD were specific to the plasma membrane of sperm cells. These two sperm specific protein dots were also found on the two-dimensional gels. These proteins may play key roles in sperm-egg recognition in the fertilization process.
Abstract (Browse 1737)  |  Full Text PDF       
A Cytological Study on the Nutrient玨Uptake Mechanism of a Saprophytic Orchid Gastrodia elata
Author: Wang He, Wang Zhen玨yu, Zhang Fu玨suo, Liu Jian and He Xiu玨xia
Journal of Integrative Plant Biology 1997 39(6)
    There are three type cells infected by the mycorrhizal fungus, ArmiUaria mellea (Vahl ex Fr. ) Karst in the tube cortex of Gastrodia elata BI., namely the passage cells, host cells and digesting cells. Ultrastructural study demonstrated that the passage cells were distroyed by the hyphae, the host cells kept symbiotic with the hyphae, but the digesting cells could become inversely parasitic on the hyphae from which nutrient were being uptaken. The detail process of the digesting cells obtaining nutrient'from the fungus is described as follows: Firstly the digesting cells began to attack the invading hyphae by releasing numerious electron-transparent vesicles of lysosomal property, secondly they took up the soluble organic material leaked out from the digested hyphae by forming many electron-dense endocytic tubes and vesicles, and finally they endocytosed and hydrolysed the insoluble hyphal walls by forming large digesting vacuole in which a piece of hyphal wall was completely enveloped.
Abstract (Browse 1756)  |  Full Text PDF       
Brassica De-Exined Pollen as a New Experimental System for Studying Pollen Germination
Author: Xu Bing-fang, Liang Shi-ping, Zhou Chang and Yang Hong-yuan
Journal of Integrative Plant Biology 1997 39(6)
    Exine deprived pollen only coated with intine has been isolated and artificially germinated in Brassica L. In view of the lack of exine and germ furrows, the de-exined pollen is an interesting experimental system for studying pollen germination. This report focuses on the microscopic and uhrastructural changes in the aspects of polarization, predetermination of the germination site and new wall synthesis during the course of isolation and germination of the de-exined pollen. It was shown that the de-exined pollen was already in an active state and had established its polarity and germination sites prior to exine detachment. The germination sites were still localized at the region of the previous germ furrows even after exine detachment. The new wall deposited at the germination sites appeared to have an important morphogenetic role in setting a limit to the size of pollen tube diameter. These results support our assumption that the de-exined pollen may have an extensive application in the research of pollen biology.
Abstract (Browse 1685)  |  Full Text PDF       
Diterpenoid Alkaloids from Aconitum vilmorinianum var. altifidum
Author: Zhang Jia-min, Wu Feng-e, Wang Ming-kui and Chen Yao-zu
Journal of Integrative Plant Biology 1997 39(6)
    A new C20-diterpenoid alkaloid, vilmorinianine, and two known alkaloids were isolated from Aconitum vilmorinianum var. altifidum W.T. Wang. The structures of these alkaloids were identified on the basis of their spectral data.
Abstract (Browse 1870)  |  Full Text PDF       


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