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Table of Content

25 August 2009, Volume 31 Issue 04

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Articles

Select AFLP Analysis of Phryma (Phrymaceae) Disjunct between Eastern Asia and Eastern North America

NIE Ze-Long^, , WEN Jun^{, ,} , SUN Hang

2009, 31(04): 289-295. doi:10.3724 SP.J.1143.2009.09075

Abstract ( 2774 )

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Although little morphological differentiation is detected between intercontinental disjunct populations of Phryma, this monotypic genus shows distinct molecular divergence corresponding to its distribution in eastern Asia and eastern North America. This study further employs amplified fragment length polymorphism (AFLP) analyses using two selective primer pairs to quantify genetic diversity and evaluate the morphological stasis . Most of the molecular variance is accounted for by variance among populations between regions .Cluster and PCA analyses revealed that Phryma constitutes twomajor groups in line with their geographic distribution, with one genetically distinct group from the eastern Asia and the other with accessions from eastern North America. The results robustly supported the distinct genetic divergence and morphological stasis in Phryma.

Select Flower Development and Cultivation of Paphiopedilum armeniacum (Orchidaceae)

PI Qiu-Xia^, , YAN Ning , HU Hong, LI Shu-Yun

2009, 31(04): 296-302. doi:10.3724 SP.J.1143.2009.09048

Abstract ( 3100 )

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The development process of the flower of Paphiopedilum armeniacum was observed . It is found that P. armeniacum developed inflorescence primordium in May , then sheath in June and July, and bracts in July and August . All
organs of the flower were formed in late August and September. After the formation of the flower , the flower organs kept on growing and developing quickly . One sterile stamen ( staminode ), two fertile stamens and the style were not integrated at the early stages of the flower development , but fused to form the gynostemium in the late phase . Because of the different relative growth rate, the relative position and morphology of these three organs were different in the late phase. In addition, several batches of P. armeniacum were introduced to Kunming and cultivated in a greenhouse from July to September, the rate of plants with flower bud had significant difference in different batches, which further verified it is a key period for P. armeniacum to form the flower organs from the late August to the early September .

Select Change of DNA Content in Male and Female Gametes of Tobacco ( Nicotiana tabacum)

LIN Mei-Zhen , DENG Hua , WEI Dong-Mei , TIAN Hui-Qiao

2009, 31(04): 303-308. doi:10.3724 SP.J.1143.2009.09035

Abstract ( 2694 )

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The nuclear DNA content of male and female gametes of tobacco ( Nicotiana tabacum) was measured using DAPI stain and microspectrofluorimetric measurement. Tobacco pollen is bicellular at anthesis, containing a vegetative cell and a generative cell , which divides to form two sperm cells in a pollen tube. The nuclear DNA content of generative cell in a pollen tube was at 2C level and that of two sperm cells in a pollen tube, which elongated in the style, at 1C level. Two sperm cells began to synthesize DNA after both were released in the degenerated synergid , and the quantity of nuclear DNA
in both sperm cells approached 2C level before both fusing with egg and central cells . During this process , the nuclear DNA content of egg cell also began to increase and approached 2C level before fusing with sperm cell . After male and female gamete fused , the nuclear DNA content of zygote reached 4C level . Therefore , the fusion of male and female gametes of tobacco was at G2 of cell cycle, and this fusion of fertilization belonged to G2 type . This result displayed the fertilization multiformity in angiosperms .

Select PLDα1 Modulated ABA-Mediated Main Root Elongation and Regulated Tip Growth of Root Hair in Arabidopsis

LI Jing, ZHANG Wen-Hua

2009, 31(04): 309-316. doi:10.3724 SP.J.1143.2009.09066

Abstract ( 3090 )

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The role of PLDα1 in ABA inhibition of main root elongation in Arabidopsis was studied . When the seedlings were treated with ABA, elongation of main root of wild- type (WT) was inhibited more obviously than that of pldα1. In main
root protoplasts, the activity of both PLDα1 and NADPH oxidase increased under ABA treatment, and the NADPH oxidase activity increased much more in WT than in pldα1. When 10μmol L^{- 1} PA was added into the growth medium, the NADPH oxidase activity increased in pldα1 protoplasts. The concentration of ROS in Arabidopsis root protoplasts increased more quickly in WT than in pldα1 after ABA addition. When H2O2 was added into the culture medium, main root elongation was inhibited in both genotypes. All these results suggested that, PA produced by PLDα1 acted as a regulator in ABAmediated
root elongation in Arabidopsis via NADPH oxidase-produced ROS. In addition, root hair of pldα1 was shorter than that of WT. On the tip of root hair, pldα1 had lower concentration of ROS and Ca²⁺ than WT .

Select Linkage Maps of the Genus Hedychium (Zingiberaceae) Based on SRAP

GAO Li-Xia , LIU Nian, HUANG Bang-Hai

2009, 31(04): 317-325. doi:10.3724 SP.J.1143.2009.09036

Abstract ( 2281 )

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In this study , using a pseudo-cross strategy , we constructed twomaps of Hedyhium , Hedychium coronarium and Hedychium forrestii based on SRAP markers . The mapping population consisted of 87 progenies, from a F1 population . A total of 414 primer pairs were screened and 92 pairs were considered . Among 398 loci , 237 loci were from the paternal parent and 161 were from the maternal parent . After χ² test and linkage analysis, two maps were constructed as following: The paternal parent contained 203 loci and spanned 1 386 . 8 cm, which spread over 23 linkage groups . The maternal parent contained 139 loci and spanned 917 . 1 cm . These loci were distributed in 18 linkage groups .

Select The Response of AtCER to Oxidative Stress in Arabidopsis thaliana

LIU Run-Hua^, , JIANG Wen-Bo ^, , YU Di-Qiu

2009, 31(04): 326-334. doi:10.3724 SP.J.1143.2009.09033

Abstract ( 2921 )

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Ceramide is a second messenger involved in many significant signal pathways of plants, such as cell growth, reduplication, differentiation, senescence and apoptosis . This work investigated the AtCER function in oxidative stress caused by H₂O₂ . The leaves of AtCER mutants cer1 , cer2 and cer3 , exhibited more severe levels of yellowing and necrotic lesions
than wild- type with H₂O₂ treatment . Further investigation by the physiological and biochemical methods, indicated that chlorophyll content of these mutants decreased more quickly than do wild type plants, and their conductivity increased significantly , and the antioxidases activity in leaves of AtCER mutants were approximately 1.5 or 3-fold higher than that inwild - type after H₂O₂ treatment . All of those results suggested that AtCER may involved in H₂O₂ -induced Oxidative stress.

Select Agrobacterium tumefaciens-mediated Transformation of Cabbage
with Betaine Aldehyde Dehydrogenase Gene

ZHOU Guo-Yan , , YANG Zheng-An , ZHANG Yin-Hua , GUO Feng-Gen ,
ZHOU Xiao-Gang , , ZHANG Shao-Song , , SUN Mao-Lin , ,
WU Shao-Yun , , DING Yue-Mei ,

2009, 31(04): 335-343. doi:10.3724 SP.J.1143.2009.09013

Abstract ( 2718 )

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Cabbage ( Brassica oleracea var. capitata) is one of the most popular and widely cultivated vegetable crops in the world . In this paper , BADH ( betaine aldehyde dehydrogenase ) gene derived from spinach was transformed into the genome of cabbage line 03079 mediated by Agrobacterium tumefaciens . To establish the ideal transformation platform, the main factors which affect the transformation efficiency were optimized through the orthogonal design of L9 (34 ) , including the type of infection medium, concentration of acetosyringone, the period of infection time and co-culture time . The results indicat-ed that No . 9 was the best treatment combination , i. e , AA liquid medium was the optimal infection medium for the cabbage transformation, and co-culturing for 2 days after infection (20min) would be favorable for the transformation . If supplemented with 200μmol L - 1 acetosyringone in infection medium, the transformation efficiency would be improved and the
transformed plant regenaration ratio reached at 54 . 26% . Based on this optimal transformtion system, we obtained many transgenic plants , and PCR analysis using BADH gene primers and Southern blot analysis indicated that the BADH gene had been integrated into genome of cabbage . The BADH enzymes activity of transgenic plants were tested after treated with NaCl, drought-tolerance and PEG stress . The results showed that the average values of the activity of BADH enzymes, which varied from 2 . 1 U to 3 . 6 U per mg in transgenic plants, were 1 to 3 times higher than that of un-transgenic plants, and there was a significant difference between transgenic and un-transgenic plants using Duncan′s Multiple Range Test. Furthermore , the average value of relative electronic conductivity ( varying from 16 . 2% to 32. 6%) of transgenic plants were significantly lower than that of un-transgenic plants , which indicated that the protection ability of membrane penetration
was enhanced when BADH activity increasing and the stress resistance of the transgenic plants was improved by introduction BADH gene into cabbage . At the same time, most of the transgenic cabbage plants were vigrously growing under drought 、salt and polyethylene glycol (PEG) stresses . The average height increasing percentage of transgenic plants were significantly higher than that of un- transgenic plants . Those results showed that the traits of drought or salt tolerance of transgenic cabbage plants were improved after transformed with BADH gene , and our transgenic plants would be used as
fundamental germplasm for stress-tolerant breeding of cabbage .

Select Analysis of Gene Expression and Source-sink Dynamics in Transportation Regulation of ABA Signal Accumulation

WEI Kai-Fa, JIA Wen-Suo

2009, 31(04): 344-352. doi:10.3724 SP.J.1143.2009.09090

Abstract ( 4385 )

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Subcellular localization of AtNCED3 , AtAAO3 and AtSDR1 proteins, and dynamic changes of ABA levels in root and leaf were investigated in the present study . Results showed that the guard cells were not the main site of ABA biosynthesis, and SDR might play an important role in ABA precursor processing and transportation . The transcription level of ABA synthase gene in roots was significantly higher than that in leaves under dehydration condition , while the ABA concentration in leaf tissues were 10 times higher than root . Detection of ABA concentration in both detached leaves and attached leaves suggested that ABA accumulation in leaves was mainly derived from root-sourced ABA, it was confirmed that ABA synthesis in roots was affected by the supply of precursor carotenoids with the fluridone treatment and shading experiments. These data revealed that ABA levels in leaves were regulated by root-sourced ABA transportation and leaf-sourced carotenoids supply. Our present results showed that vascular system might be involved in the coordination and integration of this complex regulatory mechanism for ABA signal accumulation. The conclusion provided relevant information for exploring molecular mechanism of stress signal transduction, manipulating endogenous ABA levels, and enhancing plants resistance
against environmental stresses .

Select Overexpression of miR396 miRNAs Caused Flower Stigma Curved in Arabidopsis thaliana

LIU Dong-Me^{i ,} , YANG Feng-Xi^, , YU Di-Qiu

2009, 31(04): 353-356. doi:10.3724 SP.J.1143.2009.09016

Abstract ( 2915 )

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MicroRNAs (miRNAs) are - 21 nucleotide (nt) , endogenous and encoding RNAs that play important regulatory roles in growth and development by targetingmRNAs for direct cleavage or translational repression in plants . To analyze the biological function of miR396, we placed two synthetic precursors, MIR396a or MIR396b , under the control of the enhanced
Cauliflower mosaic virus (CaMV) 35S promoter in a plant transformation vector ( pOCA30) and transformed the constructs into Arabidopsis . Genetic analysis showed that flower stigmas of 35S: MIR396 transgenic Arabidopsis were curve and siliques of miR396 transgenic plants were abnormal. In addition , Northern blot analysis showed that the miR396 miRNAs and levels of MIR396 precursor transcripts were significantly increased in transgenic miR396 plants compared with control. These results strongly supported that miR396 played an important role in plant flower development .

Select The Sample Preparation and the Improvement Method of Two-Dimensional Electrophoresis of Proteins from Antiaris toxicaria (Moraceae) Seed

YANG Jun , YANG Yun-Qiang, YANG Yong-Ping , HU Xiang-Yang

2009, 31(04): 357-362. doi:10.3724 SP.J.1143.2009.09043

Abstract ( 2323 )

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We have established the extraction method of the total proteins of Antiaris toxicaria seed and the appropriate conditions for the two-dimensional eiectrophoresis (2-DE) of Antiaris toxicaria seed proteome . Through integrating and optimizing various conditions, we also established the 2-DE system including the sample preparation method with Tris-HCL based on TCA-actone , and the first-dimensional electrophoresis with immobilized pH gradients isoelectric focusing and the second dimensional electrophoresis with vertical electrophoresis of SDS-PAGE. We have gained a good 2-DE map in the analysis of sample preparation、sample resolution、isoelectric focusing ( IEF) 、SDS-PAGE and gel staining. In order to explore the extraction method of proteome and technique of 2-DE , we compared the extraction method between the TCA- actone and the Tris-HCl , and improved the key steps of 2-DE . The Tris-HCl extraction was the most appropriate for Antiaris toxicaria seed proteome analysis because of the highest resolution and more informative 2-DE map . Those were the solid basis for the later research of studying on the discrepancy expression proteomics of Antiaris toxicaria Seed .

Select Physiological Effect of Heat Stress on Pea (Pisum sativum) Hypocotyls

TIAN Xue-Jun , TAO Hong-Zheng , LUO Jing, ZHANG Xu-Dong , TAO Fa-Qing

2009, 31(04): 363-368. doi:10.3724 SP.J.1143.2009.09017

Abstract ( 2415 )

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To explore the physiological effect of heat stress on pea ( Pisum sativum) , hypocotyl growth , membrane injury, ascorbic acid (AsA) , malondialdehyde (MDA) and the expression of Heat Shock Protein 70 (HSP70) in seedling hypocotyls of three pea varieties (produced inMengzi, Shandong and Taiwan) under different heat treatments were examined. The results showed heat stress at 48℃ resulted in decrease in germination rate of seeds and AsA content of seedling hypocotyls, and increase in membrane injury and MDA content. Longer hypocotyls of pea seedling along with higher level of AsA content, membrane thermostability and lower MDA content were found in the germinated seeds which were acclimated at 37℃ for 1 h before heat stress at 48℃ , comparing with no heat-acllimated germinated seeds (heat stres directly at 48℃ ). Except for the Taiwan pea , heat acclimation at 37℃ for 1 h couldn′t induce the expression of HSP70 in the other two varieties. However, the expression of HSP70 in all three varieties was induced by two other different heat treatments, which one was treated at 37℃ for 1 h followed by 48℃ for 2 h with recovery at 22℃ for 1 h and the other treatment was at 48℃ for 2 h . Furthermore , the expression of HSP70 of Mengzi pea was higher when suffering heat stress after heat acclimation heat stress directly .

Select Effects of Carambola ( Averrhoa carambola) Plantation on Heavy Metal Content in Soil and Heavy Mental Accumulation in Fruits

CHEN Yan , XIE Zheng-Sheng^, , ZHENG Xiao-Lin , WANG Wen-Cui

2009, 31(04): 369-373. doi:10.3724 SP.J.1143.2009.09030

Abstract ( 3024 )

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The soil heavy metal element absorption and enrichment of Carambola ( Averrhoa carambola ) were analied through the heavy metal elements content such as Cd , Pb , Cu , Hg , As, Cr of different types , different soil samples and different age trees of carambola in lianjiang green food base were investigated . The results showed that ① the soil pH≤5 . 50 which is acid soils and the heavy metal content of carambola orchard soil and fresh fruits were in conformity with the quality requirements of the production of green food; ② Heavy metal absorption carambola in the soil were different with the the different heavy metal element and different kinds ; ③ The enrichment of Cd was the most than other elements of heavy metals and the concentration coefficient was 0.947. According to the sequence as enrichment coefficients, the order was Cd (0 .947) > Hg ( 0. 098) > Cu (0. 023) > Pb (0 . 003) > As (0 . 001) = Cr (0 . 001) .

Select Plant Cyclopeptides and Possible Biosynthetic Mechanisms

DU Liang-Cheng, TAN Ning-Hua, XU Wen-Yan^, , LOU Li-Li

2009, 31(04): 374-382. doi:10.3724 SP.J.1143.2009.09093

Abstract ( 3284 )

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Plant cyclopeptides are a large group of small molecular weight natural products, typically with 4 - 10 amino acids, isolated from the leaves , stem barks , roots, and seeds of a wide variety of plant species throughout the world. The peptides are present in many Chinese medicinal plants, and their potentials have not been well exploited because of the lack of knowledge in their biosynthetic origin and mechanism. The cyclopeptides often have complex chemical structures distinct from common polypeptides ( proteins ) that are non-cyclic and gene-coded. This review discussed the potential origin of the cyclopeptides and their possible biosynthetic mechanisms .

Select A New Sesquiterpene Lactone from the Leaves of Cynara scolymus (Compositae)

LIU Rong^, , Hsieh Kun-Lung , LIU Ji-Kai

2009, 31(04): 383-385. doi:10.3724 SP.J.1143.2009.09062

Abstract ( 2672 )

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A new guaiane-type sesquiterpene lactone, named 3β, 8α, 11α, 13-tetrahydroxy-10 (14 )-guaien-1α, 4β, 5α, 6βH-6α, 12-olide (1) , together with a known sesquiterpene lactone , cynarinin A (2) , were isolated from the leaves of Cynara scolymus (Artichoke). The structure of 1 was elucidated on the basis of MS, IR, 1D, and 2D NMR .