February 1995, Volume 37 Issue 2


          Research Articles
Intrinsic GUS Activity in Various Tissues and During Pollen Development of Nicotiana tabacum
Author: Shi Hua-zhong, Yang Hong-yuan and Zhou Chang
Journal of Integrative Plant Biology 1995 37(2)
    The -glucuronidase (GUS) gene has been widely used as a reporter gene in the study of plant molecular biology and genetic engineering. One of the major reasons leading to the popularity of GUS-fusion system was the belief that there was no detectable intrinsic GUS activity in plant tissues. However, investigators have been troubled by the "false positive" results or "background" activities when GUS assays were performed. In the present experiment, histochemical observations of intrinsic GUS activity in various tissues and during pollen development of tobacco (Nicotiana tabacurn L. ) was carried out using 5-bromo-4- chloro-3-indolyl--D-glucuronic acid (X-gluc) as a substrate for overnight incubation of the treated tissues at 37. No detectable intrinsic GUS activity was found in seedling root, stem, leaf, anther wall and stigma of different stages, ovule, as well as isolated generative cell and embryo sac. During pollen development, two peaks of intrinsic GUS activity appeared, one, close to the microspore mitosis and the other from the full maturation of pollen lasting to the post-germination pollen tube stage, no or weak activity was found at other pollen developmental stages. GUS was located in the cytoplasm of the pollen. The pH value of staining solution strongly influenced the experimental results. Blue color was visualized at pH 5, even when 20% methanol or 0.2 mmol/L glucaric acid-l-4-1actone (GAL, a specific GUS inhibitor) were added. At pH 7, no detectable reaction was found at all. The aforementioned results indicate that when using tobacco pollen as the target of GUS gene transformation, the assay should be strictly controlled to neutral condition for avoiding false positive resuits.
Abstract (Browse 2187)  |  Full Text PDF       
Selection of Primary Trisomics from Anther Culture of Autotetraploid Rice (Oryza sativa L.)
Author: Liu Zong-xian and Qin Rui-zhen
Journal of Integrative Plant Biology 1995 37(2)
    The hybrids from different generations of autotetraploid rice (Oryza sativa L. ) and the original autotetraploid rice (indica and japonica) were used for anther culture, and the pollen-plantlets from them were induced. Due to the significant difference on phenotype among the trisomics and between trisomics and diploid, 15 lines of 4390 H1 induced plants were selected for chromosome study. Their PMC meiosis were observed. The results showed that the chromosomes from these plants consisted of 2n, 4n and aneuploids, and their ratios were 88.00%, 5.53% and 6.47% respectively. 272 trisomics from 284 aneuploids were identified, which acounted for 6.20% of all the pollen-plants. According to the special characters from the whole set of trisomics, they were classified as 9 types. The 9124- 7 trisomics were designated as triplo-8 by the pachytene analysis. Sowing the seeds of triplo8, the transmission rate of extra chromosome was calculated at the seedling stage of H2. The rate of trisomic was 34.11% of all plants, the agronomic characters were similar to the H1 parent plants.
Abstract (Browse 1847)  |  Full Text PDF       
Studies on Endosperm Development and Deposition of Storage Reserves in Coix lacryma-jobi
Author: Xi Xiang-yuan and Ye Bao-xing
Journal of Integrative Plant Biology 1995 37(2)
    The transition from free nuclear to cellular endosperm of Coix lacryma-jobi was eompleted 2 days after pollination. By 3 days after pollination the central cell was filled with endosperm cells. At first all cells of endosperm underwent division, later cell division was limited mainly in the peripheral region. 10 days after pollination the epidermal layer ceased its periclinal division and became the aleurone layer. Cell division persisted in the subepidermal 'cambium-like layers until the caryopsis nearly matured. Ceils of the inner region of endosperm became enlarged. Several layers of transfer cells were formed at the basal part of the endosperm. Starch grains appeared in endosperm cells on the 9th day after pollination. 10 days after pollination, lipid bodies occurred in the aleurone layer and the underlying layers. 13 and 15 days after pollination, the small vacuoles of aleurone cells contained protein and 20 days after pollenation they became aleurone grains. By 15 days after pollination pro tein bodies were formed in starch endosperm. Storage reserve deposition continued until the grain ripened. A correlation between endosperm and emoryo development was also observed.
Abstract (Browse 1748)  |  Full Text PDF       
Inhibition Effect of Zn2+ on Cytosolic Pyruvate Kinase in Passiflora edulis Leaves
Author: Guo Zhen-fei, Lu Shao-yun and Li Ming-qi
Journal of Integrative Plant Biology 1995 37(2)
    Cytosolic pyruvate kinase (PKc) activity in both mature and young leaves of Passiflora eclulis Smis was inhibited by Zn2+, and the severity of inhibition was dependant upon the concentration of Mg2+ present in the solution . Some differences in the inhibitory kinetics existed between PKc in mature and young leaves. Zn2+ showed anti-competitive inhibition of PKc in both mature and young leaves with respect to PEP and ADP, Ki (PEP) were 0. 059 mmol/L and 0. 038 mmol/L, and Ki (ADP) were 0. 074 mmol/L and 0.02 mmol/L respectively for PKc in mature and young leaves. Ki (Mg2+) of Zn2+ on PKc in both mature and young leaves were 0. 016 mmol/L and 0. 008 mmol/L respectively. These results further substantiated our previous conclusion that the in regulatory properties of PKc in mature leaves were different from those in young leaves.
Abstract (Browse 1888)  |  Full Text PDF       
Effect of Cu2+ on PS Activity and Energy Transfer of Phycobillisome in Intact Cells of Spirulina platensis
Author: Du Lin fang, Fu Hua long and Zou Xiao dong
Journal of Integrative Plant Biology 1995 37(2)
    Addition of Cu2+ at low concentrations, to intact cells of the cyanobacterium, Spirulina platensis, at room temperature, caused an enhancement in intensity of fluorescence emitted by phycocyanin and induced a blue shift at the emission peak, both of which indicated changes in energy transfer within the phycobillisomes. Cu2+ also suppressed the whole-chain electron transport activity (H2OMV) and water-splitting activity of the photosystem . When isolated phycocyanin and allophycocyanin were exposed to very low concentrations of Cu2+ ions, C-phycocyanin but not allophycocyanin, exhibited decrease not only in the absorbance in the longer wavelength (616--620 nm) region, but also in the fluorescence emission intensity at 647 nm accompanied by a blue shift to 643 nm. These results suggested that Cu2+ selectively bleach C-phycocyanin.
Abstract (Browse 1935)  |  Full Text PDF       
Isolation and Characterization of a Chinese Aneurolepidium Cell Line Resistant to Hydroxyproline
Author: Chen Hui, Kuang Bai-jian and Wang Jing-ju
Journal of Integrative Plant Biology 1995 37(2)
    A hydroxyproline (HYP) resistant cell line of Chinese aneurolepidium (Aneurolepidium chinense (Trin.) Kitag. ) was isolated under selection pressure of 20 mmol/L HYP. Comparison of the free amino acid pool levels in the cell line with that of donor showed substantial accumulation of proline (6.6 ). Enzyme examination indicated that -glutamyl kinase controlling proline biosynthesis in HR20-8 cell line had 2.5 times as much activity as that of the donor. Exogenous L-proline inhibited the enzyme activities both in the HR20-8 and the donor by the same rate of 30% at 100 mmol/L. The responses of HR20-8 cell line to NaC1, PEG and cold temperature (5 ) were also compared with those of donor and the former exhibited remarkably increased tolerance to the tested stress condition. The results showed that changes of -glutamyl kinase property exhibited the phenomenon of extra accumulation of proline which might favor the increased tolerance to NaC1, PEG and cold temperature in the resistant cell line.
Abstract (Browse 1694)  |  Full Text PDF       
A Preliminary Study on the Use of RFLP Analysis of the PCR Amplified Products in the ystematic Investigation of the Subtribe Astragalinae (Fabaceae)
Author: Ding Shi-you, Gu Hong-ya, Qu Li-jia and Chen Zhang-liang
Journal of Integrative Plant Biology 1995 37(2)
    The analysis of variation at the molecular level has been proven extremely useful in the reconstruction of plant phylogenetic relationships. A DNA fragment of 3100-base pairs (bp) in the chloroplast genes ndhF and psbA has been amplified from 9 species of 7 genera in the subtribe Astragalinae and 1 species in the subtribe Glycyrrhizinae. A proper procedure for specifically PCR-amplifing the 3.1 kb fragment was presented. By this procedure, the PCR products could be digested directly after amplification. The restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified products indicated that it had potential systematic significances in the phylogenetic studies of the subtribe Astragalinae. This approach could also reduce time, expense and the amount of DNA required, and furthermore, obtain reliable results.
Abstract (Browse 1973)  |  Full Text PDF       
Distorted Segregations of RFLP Markers and Their Distribution on Chromosomes in an indica/japonica F2 Population of Rice (Oryza sativa L.)
Author: Xu Yun-bi, Shen Zong-tan, Chen Ying and Zhu Li-huang
Journal of Integrative Plant Biology 1995 37(2)
    he segregation ratio of RFLP markers in an F2 population from indica "Zhaiyeqing 8 and japonica "Jingxi 17' of rice (Oryza sativa L., 2n= 24) was studied using 54 RFLP markers distributed on 12 chromosomes. Distorted segregation was found in 25.9% of the marker tested, which was indicated by significant deviation from the expected Mendelian segregation ratio ( I: 2: 1) at 5% or 1% level. Among the three RFLP genotypes of the F2 population Zhaiyeqing 8 genotype was significantly more than the expected, and its gene frequency was up to 52.1 %. Three positions for distorted segregation were found on chromosome 3 (RG227-RG369), 7 (RG678-RG511-RG528) and 12 (RG463-RG323). These positions could be related to gametophyte loci responsible for the distortion.
Abstract (Browse 1768)  |  Full Text PDF       
Localization of Heat Shock Protein on Cell Membranes of Phaseolus vulgaris
Author: Liu Jian, Yang Xiao-he and Wu Xian-rong
Journal of Integrative Plant Biology 1995 37(2)
    Experiment was designed to investigate the expression of heat shock proteins (HSPs) in hypocotyls of Phaseolus vulgaris L. and HSPs localization on cell membranes by means of 35S-Met labelling, fluorography of SDS-PAGE and 2-D electrophoresis. The results showed that 70 kD group HSPs were the major labelled proteins and the small HSPs accumulate largely in the fractions of plasma membrane and tonoplast membrane.
Abstract (Browse 1738)  |  Full Text PDF       
Studies on the Conversion of Chloroplast to Chromoplast During Fruit Ripening in Solanum pseudo-capsicum var. diflorum
Author: Zhu Cheng and Fang Zhao-xi
Journal of Integrative Plant Biology 1995 37(2)
    Determination of chlorophyll and carotenoid contents in the ectocarp during fruit ripening in Solanum pseudo-capsicum var. diflorurn (Veil.) Bitter revealed that the changes of fruit colour coincided with the decline of chlorophyll and the increase of carotenoid contents. The conversion of chloroplasts to chromoplasts in the fruit was studied by electron microscopy. The early green fruit was characterized by chloroplasts with a typical grana-intergranal thylakoid structure. At yellow-green fruit stage the thylakoid system was disintegrated and replaced by few non-chlorophyllous single thylakoids, with accumulation of large osmiophilic plastoglobules. The plastids developed as the so-called proplastids. These indicated dedifferentiation of chloroplasts in a ripening fruit. When the fruit reached its yellow stage, numerous large plastoglobules contained in the young chromoplasts frequently showed transitional changes to plastid tubule structure. At first, the center of plastoglobules became semi-translucent. It was believed that the young chromoplast were in an initial state of carotenoid deposition, followed by plastoglobules elongation and tubule protrution from the globules. These tubules were surrounded with an electron dense membranous sheath leaving the core semi-translucent. Concurrently a series of vesicles in different developmental stages appeared from the stroma of the plastid, likely representing a process of formation of numerous small new plastoglobules. In the chromoplasts of a ripe orange-or orange red-colored fruit only numerous tubules and small plastoglobules were present. The plastid tubules increased in number and elongated in length filling the mature chromoplast. Numerous small plastoglobules also increased and distributed in the spaces between tubules. These results indicated that the reconstruction of a mature chromoplast from a dedifferentiated plastid was really a form of redifferentiation, and it might be concluded that the conversion of chloroplast to chromoplast in the fruit of S. pseudo-capsicum var. diflorum, in fact, was a processes of dedifferentiation and redifferentiation.
Abstract (Browse 2990)  |  Full Text PDF       
Effect of Acidity and Aluminium on the Growth of Pinus massoniana Seedlings
Author: Liu Ruo-an and Liu Hou-tian
Journal of Integrative Plant Biology 1995 37(2)
    Effect of pH and A1 on the growth of masson pine (Pinus massoniana )seedlings was studied with liquid medium culture method. The masson pine seedlings exhibited strong tolerance toward acid conditions, growing normally at the pH range of 3.55.5. Growth was promoted in ultra-low concentration of Al. The masson pine seedlings were moderately sensitive to the toxicity of aluminium. The threshold of toxic concentration for aluminium to masson pine was about 4.0 ppm. Calcium ions could ameliorate the inhibition effect of A1 to some extent. The threshold ratio of A1 to Ca was about 0.2 at the level of A1 inhibition to growth of masson pine.
Abstract (Browse 1816)  |  Full Text PDF       
An Analysis of Age Structure and the Strategy for Asexual Propagation of Aneurolepidium chinense Population
Author: Yang Yun-fei, Liu Geng-chang and Zhang Bao-tian
Journal of Integrative Plant Biology 1995 37(2)
    Aneurolepidiurn chinense (Trin.) Kitag. is a rhizome grass. Among the total earing plants of the population in natural cutting grassland, 87 % was winterness plants. The oldest tillering node was four age, and its life time would reach five years. The plants of one and two age held the absolute dominant position and their proportion was 92.5% in the age structure of developing plants from the tillering nodes. The productivity of one age plants was the highest among plants of the total ages, whereas the vitalities of plants were evidently weakened by the third age. On the age structure, A. chinense population in the cutting grassland was a type of mixed population which was an expanding and a stable population. The strategy of asexual propagation, in which the age structure of the population would be regulated by developing plants from rhizome end, and the mechanism of biology and ecology on natural sustaining renovation of A. chinense grassland were comprehensively analysed.
Abstract (Browse 1855)  |  Full Text PDF       
Effects of High Temperature on the Cultures of Isolated Microspores in Brassica campestris ssp. pekinensis
Author: Liu Gong-she, Li Yan, Liu Fan and Cao Ming-qing
Journal of Integrative Plant Biology 1995 37(2)
    The influence of high temperature (33) on embryogenesis in isolated microspore culture of Chinese cabbage (Brassica campestris spp. pekinensis ) was investigated by microscpopy of FDA and DAPI. The microspores cultured at constant temperature of 25 lost their viability quickly and only few viable microspores were found after 7 days of culture. The morphology of the cultured microspores became as turgescent as the "rugby" which was similar to the mature pollen of the Chinese cabbage. The first nuclear division of the microspores was asymmetric. The microspores lost their capacity of embryogenesis under such condition. In contrast, when the microspores were cultured at 33 for 24 h before they were transfered to the culture condition of 25 , their developmental pattern was changed. Some of the microspores could remain viable even for 7 days in culture, they became rounded off. The symmetric nuclear division pattern was induced. The frequency of such division was about 40%. Of the several new cell division types observed, the symmetric type was more frequent (55%) than others. The microspores treated under 33 were able to form embryoids via embryogenesis. The critical period of high temperature treatment on microspore culture of Chinese cabbage was about the initial 12 h, if the cell division index of microspore was concerned, but the period was the initial 24 h if the frequency of embryogenesis was considered.
Abstract (Browse 1850)  |  Full Text PDF       
Karyotype and Sex Chromosomes in Cephalotaxus sinensis
Author: Chen Ke-yong
Journal of Integrative Plant Biology 1995 37(2)
    In Cephalotascus sinensis (Rehd. et Wils. ) Li the somatic chromosome number was found to be of 2n=24. Eleven pairs of chromosomes possessed their centromeres at median or median-submedian regions. The shortest pair of chromosomes was the SAT chromosome which possessed their centromeres at the submedian regions. The sex chromosomes were demonstrated by the Giemsa C-banding technique. The sexual determination mechanism of female was WZ type (2n=24=22A+WZ), and that of male was ZZ type (2n=24=22A +ZZ).
Abstract (Browse 1936)  |  Full Text PDF       
Electro-Enhancement of the Formation of Cell Clusters from Protoplasts of Tritium aestivum
Author: Dai Qun, Xia Guang-min and Guo Guang-qin
Journal of Integrative Plant Biology 1995 37(2)
    Under the treatment of 0.52.0 A direct electric currents, the number of cell clusters derived from wheat (Triticum aestivum L. ) protoplasts was dramatically increased. The effect was related to the time-length of the treatments, and appeared only when the timelength was more than the threshold. The threshold time decreased with increase of electric current intensity. The effect was not related to the starting time of the treatments in culture of the protoplasts.
Abstract (Browse 1819)  |  Full Text PDF       
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