• Title/Summary/Keyword: apoplast

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Immunological Assays of Freezing Tolerance in Barley using Antifreeze Proteins Antisera

  • Sung, Ha-Chang;Kim, Dae-In;Hwang, Cheol-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.5
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    • pp.407-412
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    • 2003
  • In order to measure an antifreezing tolerance, antifreeze proteins accumulated upon cold acclimation in apoplast were analyzed. As Dongborilho were cold-acclimated for 3 to 74 days there was an abrupt increase in apoplastic proteins up to 30 days and then decrease to the similar levels. Among the known antifreeze proteins, CLP produced in E. coli. and TLP purified from apoplast were used to generate antisera that allow to measure and localize the proteins in leaves of barley. The CLP of 27.7 kDa and TLPs of 6, 26, 27, 30, and 31 kDa were increased in their amounts in apoplast as cold treatment being longer. There was a correlation among the amounts of those proteins accumulated in apoplast and freezing tolerance as shown in field and ion leakage tests for five cultivars. The deposit of CLP was localized in the marginal area and the area adjacent to leaf vescular bundle cells in an increasing manner according to duration of cold acclimation but no variation was observed in terms of it's distribution. Based on the close correlation between levels of antifreeze proteins and degrees of freezing tolerance, the immunological methods was to develop to estimate a freezing tolerance in barley.

Variation of Antifreeze Proteins during Cold Acclimation among Winter Cereals and Their Relationship with Freezing Resistance

  • Chun, Jong-Un;Marilyn Griffith
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.3
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    • pp.172-178
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    • 1998
  • Freezing-resistant plants can survive subzero temperatures by withstanding extracellular ice formation. During cold acclimation, their leaves accumulate antifreeze proteins (AFPs) that are secreted into the apoplast and have the ability to modify the normal growth of ice crystals. Three barley, two wheat and two rye cultivars were grown under two different temperature regimes (20/16$^{\circ}C$ and 5/2$^{\circ}C$, day/night). Apoplastic proteins from winter cereals were separated by SDS-PAGE and detected with antisera to AFPs from winter rye. Apoplastic proteins accumulated to much higher levels in cold-acclimated (CA) leaves compared with nonacclimated (NA) ones in winter cereals. After cold acclimation, the protein concentration of apoplastic extracts increased significantly from 0.088 $mgmL^{-1}$ to 0.448 $mgmL^{-1}$, with about 5-fold increment. Also, the apoplastic protein content per gram leaf fresh weight in CA leaves ranged from 31 $\mu\textrm{g}$ $(gFW)^{-1}$ to 120 $\mu\textrm{g}$ $(gFW)^{-1}$ with an averaged value of 77 $\mu\textrm{g}$ $(gFW)^{-1}$, and coefficients of variation of 54.9%. The CA leaves in Musketeer (a Canadian winter rye cultivar) showed the greatest AFPs and antifreeze activity followed by 'Geurumil' (a Korean winter wheat cultivar), and 'Dongbori l' (Korean facultative barley cultivar). The proteins secreted into the wheat leaf apoplast at CA condition were more numerous than those observed in winter rye, where two $\beta$-1,3-glucanase-like proteins (GLPs), two chitinase-like proteins (CLPs) and two thaumatin-like proteins (TLPs) accumulated during cold acclimation. The proteins in barley leaf apoplast at CA conditions were a little different from those in wheat leaves. The AFPs were various among and within species. More freezing-resistant cultivars had more clear and numerous bands than less freezing-resistant ones. The high determination coefficient ($R^2$ =91 %) between freezing resistance and AFPs per gram leaf fresh weight indicated that the amount of AFPs was highly related to freezing resistance in winter cereal crops.

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Physiological and morpho-anatomical analyses of hyperhydric Arabidopsis thaliana influenced by media components

  • Nurashikin Kemat;Richard G.F. Visser;Frans A. Krens
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.255-266
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    • 2023
  • Hyperhydricity is a physiological anomaly that significantly affects the growth and proliferation rate of crops cultivated by tissue culture techniques. To better understand the mechanisms that govern hyperhydricity incidence, we examined the effects of several media components, particularly cytokinin and gelling agents. These elements were found to be influential in both in vitro propagation and the development of hyperhydricity. Our study revealed that Arabidopsis thaliana seedlings had a greater manifestation of hyperhydricity symptoms when exposed to high cytokinin concentrations compared with the control. The presence of gelrite led to the manifestation of hyperhydric symptoms by elevated water build-up in the apoplast. The phenomenon of stomata closure was observed in the hyperhydric leaves, resulting in an increased ability to retain water and a decrease in the transpiration rates when compared to their respective control leaves. Additionally, histological examinations of the cross sections of hyperhydric leaves revealed an irregular cellular arrangement and large intercellular spaces. Furthermore, hyperhydric seedlings displayed impaired cuticular development in comparison to their normal seedlings.

Antibody-Mediated Resistance to Rhizomania Disease in Sugar Beet Hairy Roots

  • Jafarzade, M.;Ramezani, M.;Hedayati, F.;Mokhtarzade, Z.;Zare, B.;Sabet, M.S.;Norouzi, P.;Malboobi, M.A.
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.692-697
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    • 2019
  • Agrobacterium rhizogenes-mediated transformation of sugar beet hairy roots expressing single-chain variable fragment (scFv) was exploited to evaluate the efficacy of four antibody-based constructs for interfering with the Beet necrotic yellow vein virus infection. The scFv specific to a major coat protein of virus, p21, was targeted to various cellular compartments including the cytosol (pIC and pICC constructs), apoplast (pIA), and mitochondrion (pIM). After mechanical virus inoculation, most of the hairy root clones expressing scFv in the cytosol displayed low virus titers while the majority of transgenic hairy root clones accumulated antibody in outer membrane of mitochondria or apoplast were infected. This hairy root system provided an efficient and rapid approach to initially investigating root disease resistance like rhizomania prior to transform whole recalcitrant plants such as sugar beet.

Silica Nanoparticles Suppress the Root Rot of Panax ginseg from Ilyonectria mors-panacis Infection by Reducing Sugar Efflux into Apoplast

  • Abbai, Ragavendran;Ahn, Jong-Chan;Mohanan, Padmanaban;Mathiyalagan, Ramya;Gokulanathan, Anandapadmanaban;Kim, Yu-Jin;Kim, Yoen-Ju;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.59-59
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    • 2018
  • Panax ginseng Meyer (Korean ginseng) is in the spotlight of Oriental medicine and is proclaimed as the king of medicinal plants owing to its adaptogenic characteristics. Ginseng root rot is a devastating disease caused by the fungus, Ilyonectria mors-panacis that generally attacks younger roots (~2 years), leading to defects in root quality, ginsenoside accumulation and also life cycle of the plant. Hence, there is an indispensable need to develop strategies resulting in tolerance against ginseng root rot. In the present study, we evaluated the effect of silica nanoparticles(N-SiO2) in Panax ginseng during I. mors-panacis infection. Long term analysis (30 dpi) revealed a striking 50% reduction in disease severity index upon 1mM and 2mM treatment of N-SiO2. However, N-SiO2 did not have any direct antifungal activity against I. mors-panacis. Membrane bound sugar efflux transporter, SWEET (Sugars Will Eventually be Exported Transporters) was identified in ginseng and as expected, its expression was suppressed upon N-SiO2 treatment in the root rot pathosystem. Furthermore, the total and reducing sugars in the apoplastic fluid clearly revealed that N-SiO2 regulates sugar efflux into apoplast. In a nut shell, N-SiO2 administration induces transcriptional reprogramming in ginseng roots, leading to regulated sugar efflux into apoplast resulting in enhanced tolerance against I. mors-panacis.

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Proteomics of plant-fungal pathogen interaction: an overview (식물과 곰팡이 병원균과의 상호작용에 대한 프로테오믹스 최근 연구 동향)

  • Kim, Jin Yeong;Lee, So Eui;Oh, Ha Ram;Choi, In Soo;Kim, Yong Chul;Kim, Sun Tae
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.1-9
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    • 2014
  • So far it has been generally considered that proteomic approaches are very useful for studying plant-microbes interaction. In this review, recent studies based on papers published from 2010 to 2013 have investigated proteomics analysis in various interaction during plant-fungal pathogen infection by means of gel-based proteomics coupled with mass spectrometry (MS)-based analysis. In rice, three papers focused on rice-Magnaporthe oryzae interaction were mainly reviewed in this study. Interestingly, another study showed proteomic changes in rice inoculated with Puccinia triticina, which is not only an fungal pathogen in wheat and but also results to the disease resistance with non-host defense manner in rice. Additionally, proteomics analysis has been widely subjected to understand defense mechanism during other crops (wheat, tomato, strawberry and mint) and their fungal pathogen interaction. Crops inoculated are analyzed to identify differentially regulated proteins at various tissues such as leaf and apoplast using 2-DE analysis coupled with various MS approaches such as MALDI-TOF MS, nESI-LC-MS/MS and MudPIT, respectively. Taken together, this review article shows that proteomics is applicable to various organisms to understand plant-fungal pathogen interaction and will contribute to provide important information for crop disease diagnosis and crop protection.

Post Genomic Approaches to Nodulation in Soybean

  • Hwang, Cheol-Ho;Lim, Chae-Woo
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.13-17
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    • 2004
  • An interaction between Legumes and Rhizobia establishes a symbiotic new organ, the nodule that supports atmospheric nitrogen fIxation. The specific communications between the microbes and legume plants are necessary for both nodulation and nitrogen fixation. Through genetic and biochemical analyses several genes playing pivotal roles in nodulation had been identified to be a receptor kinase like CALVATAl involved signal transduction for development. This emphasizes peptides as signals to be transmitted for a short or long distance transport for nodulation. In addition, a quorum sensing in rhizobia has become a focus as counterpart signal. In an attempt to reveal proteins factors and signaling molecules acting on nodulation, proteome analyses of nodule and the proteins in apoplast upon communication between Legumes and Rhizobia were performed.

A Rice Blast Fungus Alpha-N-Arabinofuranosidase B Elicits Host Defense in Rice

  • Kim, Sun-Tae
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.23-23
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    • 2015
  • Rice blast disease caused by M. oryzae is the most devastating fungal disease in rice. During the infection process, M. oryzae secretes a large number of glycosyl hydrolase (GH) proteins into the apoplast to digest host cell wall and assist fungal ingress into host tissues. In this study, we identified a novel M. oryze arabinofuranosidase B (MoAbfB) which is secreted during fungal infection. Live-cell imaging exhibited that fluorescent labeled MoAbfB was highly accumulated in fungal invasive structures such as appressorium, tips of penetration peg, biotrophic interfacial complex (BIC), as well as invasive hyphal tip. Deletion of MoAbfB mutants extended biotrophic phase followed by enhanced disease severity, whereas, over-expression of OsMoAbfB mutant induced rapid defense responses and enhanced rice resistance to M. oryzae infection. Furthermore, exogenous treatment of MoAbfB protein showed inhibition of fungal infection via priming of defense gene expression. We later found that the extract of MoAbfB degraded rice cell wall fragments could also induce host defense activation, suggesting that not MoAbfB itself but oligosaccharides (OGs) derived from MoAbfB dissolved rice cell wall elicited rice innate immunity.

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$Ca^{2+}$ Effect on the Conversion of ACC to Ethylene in Mungvean(Vigna radiata) Hypocothls (녹두(Vigna radiata)의 하배축에서 ACC의 에틸렌 전환에 미치는 $Ca^{2+}$의 효과)

  • 서효원
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.233-239
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    • 1993
  • Effect of Ca2+ on the conversion of 1-aminocycloprophane-1-carboxylic acid (ACC) to ethylene was studied with 2.5 day-old mung bean hypocotyl segments. The conversion of ACC in these tissues was inhibited by plasmolysis and sulfosuccinimidyl (hydroxyphenyl) propionate (sulfo-SHPP). The ACC induced ethylene production in HC (high calcium)-tissue grown on the Ca2+ added medium was greater than that in N (normal)-tissue. HC-tissue had a lower inhibition rate of ACC conversion by EGTA and Ca2+ -channel blockers than N-tissue. The rates of the ACC conversion by both kinds of tissues were stimulated by the Ca2+ ionophore A23187. From these results, we suggests a mechanism for the stimulative effect of Ca2+ on the conversion of ACC to ethylene as follows; in some tissues where ACC conversion is linked with plasma membrane, Ca2+ may be transported from apoplast through Ca2+ -channel into the cytoplasm ad stimulate ACC-oxidase activity.

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