Table of Content

25 October 2016, Volume 32 Issue 10

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Orginal Article

Ethylene Signaling Pathway in Regulating Plant Response to Abiotic Stress

ZHAO He, CHEN Shou-yi, ZHANG Jin-song

2016, 32(10):  1-10.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.001

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Ethylene is an important hormone which plays essential roles in regulating plant growth and development. Researches showed that ethylene was induced by different stresses and ethylene signal transduction participated in plant response to the stresses. In this review,we summarized the current advances in the role that ethylene played in regulating plant response to abiotic stresses,including drought,flooding and high salinity. Also, the potential uses and deficiencies of ethylene signaling in improving the stress tolerance of crops were discussed.

Ethylene Signaling and Its Role in Plant Stress Response

ZHANG Tao, DONG Chun-hai

2016, 32(10):  11-17.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.007

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The gaseous phytohormone ethylene affects many aspects of plant growth,development,and stress responses. In the model plant Arabidopsis thaliana,ethylene binding to receptors triggers a kinase cascade that is propagated through a number of components to the nuclear transcription factors involved in the ethylene responsive gene expression and plant responses. This article reviews the update research on regulation of the ethylene receptors,the ethylene signal transduction,and the key regulators. Meanwhile,functional roles of the ethylene signaling and regulators in plant stress responses are also discussed.

Hydrogen Sulfide Functions in Regulation of Stomatal Movement and Stress Response in Plant

CHE Yong-mei, HOU Li-xia, SUN Yan-jun, LIU Xin

2016, 32(10):  18-26.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.004

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Hydrogen sulfide（H₂S）,a recently characterized gaseous signal molecule,mediates stomatal closure induced by abscisic acid（ABA）,ethylene（ETH）etc.,and participates in plant responses to adverse environmental stresses such as salt,drought and heavy metal stresses. H₂S interacts with Ca²⁺,H₂O₂ and nitric oxide（NO）signal molecules in regulation of stomatal movement. Exogenous H₂S improves the resistance to diverse stresses by increasing antioxidant enzyme activity,up-regulating gene expression,and promoting accumulation of compatible substances such as proline and soluble sugars in plants. In this paper,the latest research progresses in the functions and molecular mechanisms of H₂S in the regulation of stomatal movement and response to diverse stresses are summarized and discussed.

A Review of Plant Cold Signal Transduction Mechanisms

HUO Chen-min, TANG Wen-qiang

2016, 32(10):  27-33.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.008

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Cold stress is one of the major environmental factors that affect the geological distribution and seasonal growth of the plants. Being exposed to low non-freezing temperature would increase plants’resistance to freezing temperature by a process called cold acclimation. During cold acclimation,plants sense the environmental low temperature,initiate cold signal transduction pathways and change the expression of genes that are critical for plants to survive the freezing temperature. In this review,we focused on recent progresses regarding our understanding of cold signal transduction mechanisms,which including the newly discovered cold signaling components CHILLING-TOLERANCE DIVERGENCE 1（COLD1）and OPEN STOMATA1（OST1）;transcriptomic studies of C-REPEAT BINDING FACTORS （CBFs）triple mutant created by CRISPR/Cas9 technology and proteomic study of early cold regulated proteins etc. Based on these results,a cold signal transduction model is summarized.

Research Progress on Drought Resistance Mechanism of Alfalfa

QUAN Wen-li, CHAN Zhu-long

2016, 32(10):  34-41.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.003

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Alfalfa（Medicago sativa）is a forage grass with the largest cultivation area in China and has important economic value and social benefits. Drought is one of the major environmental factors influencing the production and distribution of alfalfa. Soil water deficiency could lead to alfalfa’s morphological structure changes,which makes plant better adapt to the stress environment. Meanwhile,a series of physiological and biochemical responses in alfalfa plants are activated to alleviate or reduce the damage caused by drought stress. In recent years,scientists have identified drought-related genes in alfalfa and created drought tolerant transgenic plants. In this paper,alfalfa morphological,physiological and molecular mechanism in response to drought stress are reviewed and discussed. Drought stress affects phytohormone metabolism,resulting in regulation of transcription factors,which in turn modulates expression of metabolism related and stress responsive genes in alfalfa,leading to physiological and biochemical changes. Future researches should pay more attention to the regulation network in response to stresses and the excavation of new gene resources,which are beneficial to breeding programs of novel alfalfa varieties with enhanced resistance.

Progress on Mechanisms of Dehydration Tolerance in Resurrection Plants

LIU Jie, Chih-Ta Lin, DENG Xin

2016, 32(10):  42-51.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.011

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Extreme water loss is rarely found in most of higher plants,however,so-called “resurrection plants” evolved uniquely to tolerate the loss more than 90%,and resume living status rapidly after rehydration. This review summarized distribution and classification of these plants,with their morphology,physiological and molecular mechanisms in response to desiccation,and forward research and application prospects.

Progresses on Molecular Mechanisms of Low-phosphorus Tolerance in Maize

WANG Kai, LI Wen-xue

2016, 32(10):  52-57.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.002

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Maize is not only a worldwide food and feed crop but also is an important raw material for energy production and many other industrial applications. However,maize yield and quanlity are frequently threatened by phosphorus（P）limitation. The performance of high-throughput sequencing and the progress of omics in plants have shed light on the molecular machnisms of maize tolerance to low-P deficiency. In this review,we concentrates the present progresses of the molecular mechanisms of low-P tolerace in maize,especially on P deficiency induced genes,omics and QTL mapping. This would be useful for improving low-P tolerance of maize varieties.

Plant Phospholipase D Signaling Pathway in Response to Salt Stress

WANG Pei-pei, SONG Ping, ZHANG Qun

2016, 32(10):  58-65.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.012

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Soil salinity is a global ecological problem,which has a great negative impact on ecological environment and agricultural production. Researches showed that Phospholipase D（PLD）was a key enzyme in catalyzing lipids and response to abiotic stresses. PLDs contained diverse structures,distinguishable biochemical and regulatory properties,which produced phosphatidic acid（PA）and participated in plant response to various stresses. This review summarized the current advances in regulating plant salt tolerance,and discussed the putative molecular mechanisms of PLD in sensing and response to salt stress,which might benefit the study of plant salt resistance and crop genetic enhancement.

WRKY Transcription Factors in Regulation of Stress Response in Plant

SUN Shu-hao, YU Di-qiu

2016, 32(10):  66-76.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.009

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WRKY transcription factor families are characterized by a highly conserved WRKYGQK domain and involved in plant development,metabolism,answering to comprehensive biotic or abiotic stress. Recently,the research of WRKY transcription factors concentrate on stress response signaling network in different species. It reviewed progress of WRKYs members,and indicated that WRKY transcription factors play a heavy role in plant growth and regulating stress response. At the same time,there is less reported of WRKYs function in plant species besides model plant Arabidopsis thaliana and most of them focus on systematic research and analysis. In addition,numerous networks of WRKY transcription factors are still unclear.

Function Exploration of Grape WRKY Family Proteins Under Abiotic Stresses

ZHU Dan, MA Qian, HAO Jie, LIU Xin

2016, 32(10):  77-83.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.006

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Abiotic stresses caused by drought,salt and cold restrict the development of grape industry. WRKY,as a large transcriptional regulation factor gene family,plays important role in multiple signal-transduction pathways. With the advances in high throughput sequencing technology and a variety of research techniques,more and more WRKY protein’s function and mechanism in abiotic stresses have been validated,which establish a theoretical basis and provide a rich candidate gene resources for grape abiotic stress resistant mechanism research and screening excellent varieties through molecular breeding. This review will summarize the identification and classification,drought,salt and cold stresses response mechanism of grape WRKY family proteins in detail.

Endoplasmic Reticulum Stress Response in Plants

YANG Zheng-ting, LIU Jian-xiang

2016, 32(10):  84-96.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.005

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Endoplasmic reticulum（ER）is an important organelle in eukaryotic cells where all the secreted proteins and most of the membrane protein are synthesized. When plants are exposed to various stresses,such as heat and high salinity,unfolded and misfolded proteins are accumulated in the endoplasmic reticulum（ER）,which triggers a well-conserved pathway called the unfolded protein response （UPR）to mitigate the ER stress and promote cell survival. In this review,we summarized the current advances in the understanding of the membrane-associated transcription factors and ER stress responses in plants. We also discussed future directions in this field and the potential uses of the ER stress signaling pathway for improving the stress tolerance of plants.

Recent Progress in Molecular Mechanism of Rice Disease Resistance

LI Zhi-qiang,WANG Guo-liang,LIU Wen-de

2016, 32(10):  97-108.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.010

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Rice as one of the most important crop which is the staple food for more than half of the population in the world. Maintaining high and stable rice yield are closely related to the national economic development. However,various plant diseases and insect pests pose to seriously yield production lost threat on rice. Breeding and plant high disease resistance cultivar is the most economic effective,safe,healthy and environment friendly rice disease breeding strategy,and in-depth studies in rice disease resistance molecular mechanism can provide important theoretical basis for cultivation of high disease resistance rice varieties. In the past two decades,scientists have made important progress in the molecular mechanism of rice disease resistance. This review summarizes the recent progress towards understanding the recognition and signaling events that govern rice innate immunity and the application in rice disease resistance breeding. We also discuss and prospect the challenges and future directions for research that will further our understanding of rice disease resistance.

Research Advances in the Mechanism and Signal Transduction of Plant Disease Resistance

DING Li-na, YANG Guo-xing

2016, 32(10):  109-117.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.013

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The mechanism of plant disease resistance is the focus of plant pathology. With the development of the molecular biology,people has gained more and more insights into the interaction between plant host and pathogens. In this review,the recent research advances on molecular mechanism of plants disease resistance are summarized. It also presented the function of several signal molecular such as calcium Ion,nitric oxide,reactive oxygen species,salicylic acid,jasmonic acid/ethylene,and heterotrimeric G proteins,which are known to play important role in inducing plant defense response. In addition,the prospect of future work on plant defense research is discussed in the review, aiming to offer new thinking for the development of disease control stratey.

Research Progress of Stress Tolerance and Breeding Research on Lycium ruthenicum Murr

LIU Li-ping, ZHANG Dong-zhi, ZHANG Chong, CHEN Jin-huan

2016, 32(10):  118-127.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.014

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Excellent-strains which have high abiotic stress resistance are eagerly required in order to improve and utilize the broad saline-alkaline land,as well as solving the ecological problems and bringing some economic benefits. As one of the wild shrub species covering functions such as saline alkali land green,economic and medicinal values,Lycium ruthenicum Murr have a great application prospect to be used as an ecological spcies or an economic forest tree species. Here,we summarize stress resistance study and the basic research about breeding of this species in recent years,aiming at providing the reference for Lycium ruthenicum Murr resistance evaluation and the job of breeding.

Advances in Studies on the Acclimation of Antarctic Ice Microalgae to Extreme Environments

WANG Yi-bin, ZHANG Ai-jun, LIU Fang-ming, ZHENG Zhou, MIAO Jin-lai

2016, 32(10):  128-134.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.015

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Antarctic ice microalgae are the phytoplankton or microalgae that thriving in the pelagic,ice and meltwater environments of the Antarctic,which are the main sources of primary production in an otherwise barren region. Antarctic microalgae possess unique adaptations that allow them to proliferate in extreme conditions characterized by low or freezing temperature,seasonal light,strong ultraviolet radiation and high salinity fluctuations. Thus,understanding the underlying mechanisms of their acclimation has gained strong interests in the past years. This paper then aims to provide a succinct review on the progress,status and new paradigms of research on Antarctic microalgae. These new findings on Antarctic ice microalgae will help us gain deeper insights into the general adaptive mechanisms of all phytoplankton to extreme environment and the potential role of these organisms in the important biogeochemical cycles.

The Influence for Two Corps Physiological Index by the Different Petroleum Density

LUO Xiao-fang, CHEN Li-hua, WANG Dong-mei, CHEN Fang-yan

2016, 32(10):  135-140.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.016

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The experiment have been selected the Triticum aestivum Linnand Zea mays L. two different plants for 60 days imitate in the laboratory. According to measure the oil contaminated soil toleranceof two different plants and the oil pollution stress on petroleum contaminated soil restoration potential effect,which hope to provide a theory basis in future remediation cases. The experiment set up 0%,0.5%,1.5%,3% four groups of polluted soil respectively,and doing the experiment that culture the tested plants of wheat and corn in the greenhouse by pot. The results shows that the germination rate of tested plants in the petroleum coercion all shows downward trend,and have an extremely influence on the result when the density reach to 3%;The petroleum density to the wheat and corn have an inhibition actions,compare CK with the height,the number of leafs,the dry substance,which have an obvious descend trend. The content of chlorophyll decreased with the petroleum density increasing for wheat and corn. The content of MDA,soluble protein in the wheat leafs,which shows firstly increased and then decreased with the petroleum density increasing,but the content of MDA increasing with the petroleum density increasing,the content of soluble protein shows a decreased trend with the petroleum density increasing.

Effect of Iron Deficiency on Physiological Characteristics in Seedlings of Adzuki Beans（Vigna angularis）

WANG Jin-long, WANG Wei-wei, WAN Ping, YANG Kai, ZHAO Bo, LI Yi-song

2016, 32(10):  141-147.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.017

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To investigate the effects of iron deficiency on physiological characteristics in seedlings of Adzuki Beans（Vigna angularis）,four accessions of Adzuki Beans（13MY001,13MY007,FMY020 and JN6）were cultured in solution with or without Fe（II）-EDTA. Some traits were analyzed in 7,14,and 21 days. The results show that the iron deficiency significantly reduces the chlorophyll content,photosynthetic rate,root surface area,total root length and total biomass,and increase the Fe（Ⅲ）reductase activity and root respiration rate of Adzuki Beans seedlings. Furthermore,the differences between different cultivars are significant. The results obtained in this experiment indicated that the combination of the chlorophyll content,photosynthetic rate,and Fe（Ⅲ）reductase activity and root respiration rate could be a promising screning index for selecting of Fe-efficient genotypes in Adzuki Beans．

Cloning and Analysis of Structure Variation and Expression of MeNCED3 Gene in Cassava

DING Ze-hong, FU Li-li, TIE Wei-wei, YAN Yan, HU Wei

2016, 32(10):  148-153.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.018

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To reveal the roles of MeNCED3 gene in abiotic stresses（e.g.,drought）in cassava. RT-PCR method was applied to clone MeNCED3 gene from cassava leaves,MEGA software was used to construct its phylogenetic tree,DnaSP software was used to analysis its structural variation,and quantitative RT-PCR（qRT-PCR）was used to explore its expression patterns in response to different abiotic stresses. Results showed that a NCED（9-cis-epoxycarotenoid dioxygenase）gene,MeNCED3,was cloned from cassava. This gene had a 1 803 bp open reading frame,encoded 600 amino acids,and contained a NCED conserved domain. Phylogenetic analysis showed that MeNCED3 had close genetic relationship with its homologs from Populus trichocarpa and Salix purpurea,and the sequence similarity was up to 83.9% and 82.5%,respectively. Genetic structural variation revealed that a total of eight mis-sense mutations were identified between wild and cultivated cassava species,of which five may be associated with the different expression levels of MeNCED3. qRT-PCR analysis revealed that MeNCED3 expression was higher in roots than in leaves and stems. In addition,the expression of MeNCED3 was significantly induced by PEG,ABA and NaCl treatments. MeNCED3 played a regulatory role in both salt and osmotic stresses at the transcriptional level and it could be served as a candidate to further study its functions in drought resistance in cassava.

Cloning and Stress Response Analysis of PwRbcS in Picea wilsonii

JU Dan, HU An-ni, ZHANG Jie, ZHANG Ling-yun

2016, 32(10):  154-162.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.019

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RbcS gene encodes a subunit of nuclear plant photosynthesis 1,5 two ribulose bisphosphate carboxylase / oxygenase. In this study,PwRbcS cDNA was obtained using RACE -PCR method. Bioinformatics analysis and real-time PCR technique were used to determine the relative expression of PwRbcS. The results showed that the full-length of PwRbcS cDNA was 936 bp,including the ORF of 552 bp encoding 186 amino acids. The theoretical molecular weight of PwRbcS is 20.712 6 kD and the pI value was 9.07. RT-qPCR showed that PwRbcS is mainly expressed in the needles,especially in mature needles. The expression of PwRbcS changed obviously under the treatment of NaCl,ABA and low temperature,respectively. After NaCl treatment,the expression level of PwRbcS changed significantly,reaching to 14 times than the control at 6 h. The expression level of PwRbcS reached to 24 times than the control after 8 h-ABA-treatment. And under low temperature stress,the expression increased gradually to 12 times than the control and followed by a drop,while under high temperature and PEG stress expression level did not change significantly. Therefore,PwRbcS may play an important role during abiotic stresses in planta.

Cloning of Small Heat-shock Protein（HSP）Gene from Sugarcane and Analysis of Its Expression Under Drought Stress

LIANG Pan-xia, HUANG Xing, LI Yang-rui

2016, 32(10):  163-169.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.020

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The present experiment was conducted to clone Small Heat-shock Protein（sHSP）in sugarcane to explore mechanism of stress resistance in sugarcane. sHSP was amplified using RT-PCR from sugarcane leaves,the characteristics of the deduced protein were analyzed using bioinformatics software and its expression was analyzed using quantitative real-time PCR. The results showed that the cDNA of HSP gene was 659 bp in full length with a 459 bp open reading frame,and encodes a putative sHSP protein with 152 amino acids. Comparison of the amino acids sequences homology in sHSP protein from 14 different species indicated that,the sHSP protein had 69% to 96% identity in amino acids sequence with other plants. The deduced amino acids sequence not only contained a typical sHSP domain,but also was conservative. The results of quantitative real-time PCR analysis showed that the mRNA of sHSP was decreased initially and then increased with time under drought stress. These results suggested that sHSP might be involved in function of water stress and drought resistance in sugarcane enhanced by Si application.

Sequence Divergence and Analysis of Expression Difference of Vacuolar Invertase Gene EhvINV from Different Resistant Populations in Elsholtzia haichowensis

CAI Shen-wen, XU Zhong-rui, XIONG Zhi-ting, WANG Jia-zhen, CHEN Yao

2016, 32(10):  170-179.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.021

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Specific primers were designed to clone the cDNA sequences according to the EhNvINV（JX500755）and EhCvINV（JX500756）from GenBank. The DNA and deduced amino acid sequences were analyzed by bioinformatics methods. The three-dimensional structures were constructed by homologous modeling. The structures of vacuolar invertase from two populations of E. haichowensis and non-tolerant population with each single point mutation in complex with sucrose were simulated by AutoDock 4.0. Transcript expression of EhNvINVand EhCvINV under copper tress was analyzed by real-time PCR. The results showed that there were two divergent amino acids at position 114 and 346 between EhNvINV and EhCcINV. The three-dimensional structures were exactly similar between EhNvINV and EhCvINV. It showed difference at Glu114/Gln114 and Leu346/Pro346,which were divergent sites. The structures of catalytic active center of EhNvINV,EhCvINV,and simulated mutants（EhNvINV-E114Q,EhNvINV-L346P）binding with sucrose showed no significant differences. However,there were differences on spatial position. The result of real-time PCR indicated that the transcript expression of EhCvINV induced by copper stress after 7 days,however,the transcript expression of EhNvINV inhibited by copper stress after 7 days.

Insect Resistance Analysis of an Agglutinin Gene PPA2 Cloned from Pinellia pedatisecta

LIU Ding, CHEN Jin, LIU Zhi, ZHU Sheng-wei

2016, 32(10):  180-187.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.022

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Pinellia pedatisecta Agglutinin（PPA）belonging to mannose-binding lectin muti-family has insect resistant activity. A new gene,named PPA2,encoding a PPA protein was cloned by RT-PCR from young leaves of pinellia pedatisecta. The PPA2 consists of an ORF of 408 bp coding 135 amino acids（14.7 kD）,and contains a mannose conservative domain with three mannose-binding sites. The transgenic tobacco plants mediated by Agrobacterium with leaf disc method were obtained under the control of CaMV 35S promoter and Arabidopsis thaliana phloem promoter AtPP2. A few of homozygotic transgenic plants with single copy insertion and high expression level were indentified in T3 generation,and then these plants were divided into high,middle,and low classes according to the level of expression protein detected by western-blot analysis. The results of aphid bioassay showed that all transgenic plants had obviously insecticidal activity against the tobacco aphids（Myzus nicotianae）,and the CaMV 35S-PPA2 transgenic plants had the highest insecticidal activity,while the aphid inhibition was much lower in AtPP2-PPA2 transgenic plants than in CaMV 35S-PPA2 ones. These findings suggest that PPA2 is a suitable candidate gene for high efficient aphid resistance and has important application prospect.

Roles of Nicotiana tobacum NtWRKY40 in Plant Responding to Virus Infection

LIU Jing-jing, CHENG Chun-ling, XI Yu-zhen, WEI Shu

2016, 32(10):  188-198.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.023

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Some of WRKY transcription factors have involved in plant abiotic defense. A WRKY gene,a homologue of Arabidopsis WRKY40,was found being dramatically induced by plant virus and further characterized in this study. To verify its function,NtWRKY40over-expressed tobacco plants were established. After virus infection,the transcription levels of NtWRKY40 and virus related genes were quantified using quantitative real time PCR at three stages（6 h,3 d,9 d after infection）. Results showed that tobacco mosaic virus infection led to 150 fold higher of NtWRKY40 transcript in non-transgenic wild type tobacco than non-infected control. No significant change in NtWRKY40 transcription levels was noted in the NtWRKY40 over-expressed tobacco after virus infection.NtWRKY40 over-expressed tobaccos were more sensitive to TMV infection than non-transgenic wild type. The data suggested that NtWRKY40 was a negative regulator in responding to TMV. In addition,an artificial miRNA vector amiR167-PVY was constructed,for silencing the coat protein gene of Potato virus Y,using the miR167 precursor of Nicotiana tabacum as backbone and cauliflower mosaic virus 35S promoter. Over-expression of amiR167-PVY exhibited inhibitory effect on the virus. The results provide the basis for the further research on amiRNA-mediated virus resistance mechanism,and contributes to antiviral transgenic plants cultivation with the optimal target sequence.

The Expression Differences Between Bacterial Blight Resistance Genes and Signal Materials in Embryonic Cell and Leaf of O.meyeriana Bail

LIU Lin, PENG Bo, JIANG Chun-miao, CHENG Zai-quan

2016, 32(10):  199-204.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.024

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In order to research the effect of the Oryzae on the resistance signal substance and resistant bacterial blight gene of O.meyeriana Bai,the O.meyeriana embryonic cells（callus,specialized cells）and leaf cells（no-specialized cells）were treated with the Oryzae. Then the content change of resistance signal substance salicylic acid（SA）and jasmonic acid（JA）,and the expression of genes related to bacterial blight resistance were detected. The research has shown that the content of SA and JA in callus before and after treatment was higher than that in leaf cells. This may be related to the cell wall of no-specialized callus which are thinner in order to prevent the invasion of pathogenic bacteria.In order to prevent the invasion of pathogenic bacteria,the expression was increased. At the same time,it shows that the ability to resist the Oryzae of the O.meyeriana is in the primary germ cell. The ME207 gene was expressed before and after the specialized differentiation of the O.meyeriana cells. But the ME281 gene is an inducible expression gene before the specialized differentiation of the O.meyeriana cells,which is a constitutive expression gene after the specialized differentiation of the O.meyeriana cells.

Evaluation of Candidate Reference Genes for Normalization of Quantitative RT-PCR in Cinnamomum camphora Under Various Abiotic Stresses

ZHANG Jia-jia, ZHANG Li-wei, LI Yong-peng, JIAO Xiao-lin, ZHU Feng-ling, DU Li

2016, 32(10):  205-211.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.025

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This study is to obtain the optimal reference genes of Cinnamomum camphora under different abiotic stresses. The expression level and expression stability of two candidate reference genes CcACTc and CcEF1α in C. camphora under various abiotic stresses were analyzed using quantitative real-time polymerase chain reaction（qRT-PCR）technique and BestKeeper software,respectively. The results indicated that,CcEF1α was relatively stable under low temperature treatment,and performed higher stability under other abiotic stress,such as drought,high salinity and ABA treatment compared to CcEF1α. Accordingly,CcEF1α is a considerable reference gene for quantitative analysis of stress-related genes in C. camphora.

Drought-tolerance Analysis and SRAP-based Germplasm Identification of Chinese Fir Clones

SU Yan, ZHANG Yi-chi, HU De-huo, ZHENG Hui-quan

2016, 32(10):  212-218.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.026

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It was to evaluate the drought-tolerance of the elite clones（T-c01,T-c04,T-c07,T-c08,T-c22,T-cF1）of Chinese fir（Cunninghamia lanceolata（Lamb.）Hook）,and to generate clone-specific digital DNA fingerprinting ID,aiming to provide basis for the application and protection of these clones. Different plant physiological and biochemistry indexes including relative water content,malondialdehyde and proline content,and activity of superoxide dismutase,peroxidase and catalase,were measured for the present clones under PEG-6000 simulated drought condition. The results indicated that there had comprehensive change in physiological and biochemistry indexes among clones,while different clone seemed to have rather different systematic drought-response manner. According to the subordinate function values,a sequenced drought-tolerant order was identified,showing as T-c01 > T-c22 > T-cF1 > T-c07 > T-c04 > T-c08. For sequence-related amplified polymorphism（SRAP）based-germplasm discrimination assay,16 SRAP primer combinations produced 131 clear and stable bands with a percentage of polymorphic bands of 66.8%,allowing a generation of clone-specific digital DNA fingerprinting ID by primer set of Me1/Em1+Me4/Em21+Me11/Em5 with confidence probability of 99.99998%.

Screening,Identification and Fermentation Condition Optimun of a Siderophore-producing Bacteria WN-H3 from Rhizosphere of Ryegrass

CHEN Wei, SHU Jian-hong, CHEN Ying, ZENG Qing-fei, WANG Xiao-li, LU Rui-xia, FU Wei

2016, 32(10):  219-226.  doi:10.13560/j.cnki.biotech.bull.1985.2016.10.027

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By mean of the chrome azural S（CAS）assay,a high-affinity siderophore- producing bacteria WN-H3 was screened out from 28 bacteria,which were isolated from the ryegrass（Lolium perenne L.）rhizosphere soil. According to morphological,biochemical and physiological characteristics,16S rDNA sequence and phylogenetic analysis,the strain was identified as Pseudomonas vranovensis. The optimal medium components（carbon and nitrogen sources）and growth conditions（temperature,rotation speed and initial pH）for siderphore production by strain WN-H3 were researched by one variable-at-a-time method. The result demonstrated that high siderophore production was yeilded at high siderophore- producing ability of 80.4% under the condition of 10 g/L sucrose,5 g/L yeast extracion,5 g/L NaCl,temperature 28℃,initial pH7.5,180 r/min.