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Cassava (Manihot esculenta Crantz) is the most important staple food for more than 300 million people in Africa, and anthracnose disease caused by Colletotrichum gloeosporioides f. sp. manihotis is the most destructive fungal disease affecting cassava production in sub-Saharan Africa. The main objective of this study was to improve anthracnose resi...
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... leaves of rice (Oryza sativa) cultivar Japonica were collected from Mwea Rice Irrigation Scheme fields in Kenya and used for isolation of Ostlp gene. RNA was extracted from 100 g of leaf tissue using RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). First-strand cDNA was synthesized from DNAse treated RNA using RevertAid First Strand cDNA synthesis Kit (Thermo Scientific, USA). The full-length cDNA of Ostlp (Os12g0628600) was polymerase chain reaction (PCR) amplified using forward primer, 5 0 - CCATGGCGTCTCCGGCCACCTCTTCCGCT-3 0 and reverse primer, 5 0 - CACGTGTTATGGGCAGAAGACGACTTGGTA-3 0 , containing NcoI and PmlI sites (underlined) at their respective 5 0 -ends. The PCR prod- ucts were cloned into the pJET vector (Fermentas). The vector was digested with NcoI and PmlI and inserted into the pCAM- BIA1305 vector (CAMBIA, Canberra, Australia) replacing the GUS gene. The hygromycin phosphotransferase gene (hpt) in pCAM- BIA1305 was substituted with neomycin phosphotransferase (nptII) gene. The resulting recombinant PCam:Ostlp vector contains rice tlp and nptII genes driven by the CaMV35S promoter (Fig. 1). The pCam:Ostlp plasmid was transferred to Escherichia coli DH5a strains by heat shock method. Transformants were selected on LB agar plates containing 50 mg/l kanamycin and confirmed by restriction digestion with NcoI and PmlI and sequencing. The pCam:Ostlp was mobilized into Agrobacterium tumefaciens strain LBA4404 by electroporation (Gene pulser Ò II, Bio-Rad Laboratories Inc., Richmond, CA). The clones on the LB plate with kanamycin, rifampicin and streptomycin were confirmed by PCR with primers specific to full-length Ostlp. The Agrobacterium strain LBA4404 har- bouring pCam:Ostlp was maintained on LB medium (supplemented with 50 mg/l rifampicin, 50 mg/l kanamycin and streptomycin 100 mg/l) and used for transformation ...
Citations
... A recent study showed that 19 classes of PR proteins can be distinguished based on structural similarity and functional activity [1,4], of which, thaumatin-like proteins (TLPs)-the homologies of sweet-tasting thaumatin isolated from the plant Thaumatococcus daniellii-belong to the PR-5 family [5,6]. Growing evidence shows that TLPs are involved in resistance to a variety of fungal diseases in many species, such as Gossypium hirsutum [7], Gossypium barbadense [8], Solanum lycopersicum [4,9], Phyllostachys edulis [10], Ganoderma lingzhi [11], Triticum aestivum L. [12,13], Carya cathayensis [14], Fragaria ananassa [15], Allium sativum L. [16], Pinus radiata [17], Lentinula edodes [18], Camellia sinensis [19], Cucumis melo L. [20], Avena nuda [5], Musa acuminate [21], Manihot esculenta [22], Vitis amurensis [23], Populus szechuanica [24], Camellia sinensis [25], ...
Thaumatin-like proteins (TLPs) comprise a complex and evolutionarily conserved protein family that participates in host defense and several developmental processes in plants, fungi, and animals. Importantly, TLPs are plant host defense proteins that belong to pathogenesis-related family 5 (PR-5), and growing evidence has demonstrated that they are involved in resistance to a variety of fungal diseases in many crop plants, particularly legumes. Nonetheless, the roles and underlying mechanisms of the TLP family in legumes remain unclear. The present review summarizes recent advances related to the classification, structure, and host resistance of legume TLPs to biotic and abiotic stresses; analyzes and predicts possible protein–protein interactions; and presents their roles in phytohormone response, root nodule formation, and symbiosis. The characteristics of TLPs provide them with broad prospects for plant breeding and other uses. Searching for legume TLP genetic resources and functional genes, and further research on their precise function mechanisms are necessary.
... DC3000, Puccinia triticina, and Colletotrichum gloeosporioides f. sp. manitiss [7,8,33,34]. In addition, TLPs can also be induced by bacterial pathogens, abiotic stresses (e.g., wounding, drought, osmotic stress, low temperature, high salt, and UV radiation), and phytohormones [35][36][37]. ...
Although thaumatin-like proteins (TLPs) are involved in resistance to a variety of fungal diseases, whether the TLP5 and TLP6 genes in tomato plants (Solanum lycopersicum) confer resistance to the pathogenesis of soil-borne diseases has not been demonstrated. In this study, five soil-borne diseases (fungal pathogens: Fusarium solani, Fusarium oxysporum, and Verticillium dahliae; bacterial pathogens: Clavibacter michiganense subsp. michiganense and Ralstonia solanacearum) were used to infect susceptible “No. 5” and disease-resistant “S-55” tomato cultivars. We found that SlTLP5 and SlTLP6 transcript levels were higher in susceptible cultivars treated with the three fungal pathogens than in those treated with the two bacterial pathogens and that transcript levels varied depending on the pathogen. Moreover, the SlTLP5 and SlTLP6 transcript levels were much higher in disease-resistant cultivars than in disease-susceptible cultivars, and the SlTLP5 and SlTLP6 transcript levels were higher in cultivars treated with the same fungal pathogen than in those treated with bacterial pathogens. SlTLP6 transcript levels were higher than SlTLP5. SlTLP5 and SlTLP6 overexpression and gene-edited transgenic mutants were generated in both susceptible and resistant cultivars. Overexpression and knockout increased and decreased resistance to the five diseases, respectively. Transgenic plants overexpressing SlTLP5 and SlTLP6 inhibited the activities of peroxidase (POD), superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) after inoculation with fungal pathogens, and the activities of POD, SOD, and APX were similar to those of fungi after infection with bacterial pathogens. The activities of CAT were increased, and the activity of β-1,3-glucanase was increased in both the fungal and bacterial treatments. Overexpressed plants were more resistant than the control plants. After SlTLP5 and SlTLP6 knockout plants were inoculated, POD, SOD, and APX had no significant changes, but CAT activity increased and decreased significantly after the fungal and bacterial treatments, contrary to overexpression. The activity of β-1,3-glucanase decreased in the treatment of the five pathogens, and the knocked-out plants were more susceptible to disease than the control. In summary, this study contributes to the further understanding of TLP disease resistance mechanisms in tomato plants.
... Several studies reported that over-expression of TLP in plants leads to improved tolerance against various fungal pathogens ( Table 2). Ojola et al. [61] tested the anti-fungal activity of transgenic cassava (Manihot esculenta Crantz) overexpressing rice TLP (OsTLP) against Colletotrichum gloeosporioides. Using leaves and stem cutting bioassays, severe necrotic symptoms were observed in the nontransgenic control as early as 2 days post-inoculation. ...
... Using leaves and stem cutting bioassays, severe necrotic symptoms were observed in the nontransgenic control as early as 2 days post-inoculation. In contrast, transgenic cassava displayed delayed disease symptoms which appeared 6 days-post-inoculation [61]. Similarly, Yan et al. [7] integrated another TLP gene, TLP29, from wild grape Vitis quinquangularis cv. ...
Unrelenting threats of fungal diseases have caused huge losses to farmers around the
world. Despite efforts and progress made in strengthening the conventional breeding
strategies, a sustainable solution is yet to be discovered. Thus, the development of
transgenic crops using anti-fungal genes such as thaumatin-like protein (TLP), a
member of class 5 pathogenesis-related protein (PR-5), is deemed as a viable solution
to achieve better resistance trait. However, the plant genome usually has multiple TLP
copies with varying signature domains and motifs. It is possible that not all of the copies
exert anti-fungal activity. In addition, the biological roles of TLPs underlying their
ability to improve the plants’ tolerance are still inconclusive. This mini review will
discuss the unique characteristics of TLPs, which might contribute to their anti-fungal
property as well as the various biological roles the proteins may play in the plants’
defense mechanism against fungal infection. Several TLPs that have been proven to
possess anti-fungal activity will also be discussed. Moreover, the impact of transgenic
plants overexpressing TLPs in combating fungal diseases will also be elaborated via
several successful transgenic researches. The information presented in this mini-review
will greatly highlight the potential of TLPs as an anti-fungal agent, especially in the
generation of transgenic plants with improved tolerance against fungal diseases.
... Cassava (Manihot esculenta Crantz), a perennial shrub of the Euphorbiaceae family, is widely cultivated in tropical and subtropical regions. It is a root crop that is a staple food for approximately 800 million people worldwide [1,2]. It is also an important raw material for the production of starch, processed food, and biofuels [3,4]. ...
Cassava is an important crop for food, feed, and industrial raw materials. Given that traditional conventional breeding is restricted by various factors, biotechnology breeding has become an important breeding method. Tissue culture regeneration is the basis of biotechnology breeding. This chapter reviews the establishment and development of cassava tissue culture and regeneration systems and the technical processes of tissue culture and regeneration starting from the induction of explants of tissue-cultured cassava plantlets to embryogenic calli, isolation to protoplasts, culture to embryogenic calli followed by differentiation into embryos, and then sprouting, stemming, and rooting into complete plants. This chapter focuses on the technical processes from protoplast to complete plant and summarizes the important influencing factors of protoplast regeneration, which is the key and difficult point in the entire regeneration process of cassava protoplasts. This chapter aims to provide technical guidance for cassava protoplast regeneration, offer useful inspiration and reference for cassava tissue culture, and lay a foundation for the genetic improvement of cassava.
... On the contrary, PaGAPDH, PaHSP70 and PaThau showed elevated expression in the OPWV-treated drought-stressed samples. In addition, several studies showed that GAPDH, HSP70 and Thau play an essential role in plant biotic and abiotic tolerances [41][42][43][44]. These findings suggest that OPWV could modulate these drought-responsive genes to improve plant drought tolerance. ...
Drought stress severely threatens plant growth, yield and survivability. Wood vinegar, formed by the condensation of smoke produced during biochar production, has been shown to promote plant growth and enhance stress tolerance. They have now been recognized as a sustainable alternative and are frequently used exogenously to support plants coping with environmental stress. This study aimed to evaluate the efficacy of oil palm wood vinegar (OPWV) in mitigating the adverse effects of drought stress on Pandanus amaryllifolius. The optimal concentrations and frequencies of OPWV application were determined before the drought treatment. The results showed that the imposed drought stress negatively affected the plant growth parameters but applying OPWV at 1:500 dilution at 3-day intervals for 12 days increased its tolerance. These include increased leaf relative water content, root-to-shoot ratio, relative stem circumference, chlorophyll pigments and antioxidant enzyme activities. In contrast, the drought-stressed plants treated with OPWV showed decreased relative electrolyte leakage, hydrogen peroxide, proline, malondialdehyde, and enhanced drought-responsive gene expressions, such as HSP70, GAPDH, and Thau, while ENO and β-Fruc were reduced. These biostimulatory effects of OPWV might be due to several antioxidant compounds, such as anthranilic acid, tetrasiloxane, syringol, guaiacol, and catechol. Altogether, our results showed the effectiveness of OPWV in alleviating the adverse effects of drought stress, and as such, OPWV could be potentially applied in agriculture.
... Colletotrichum species are among the top 10 plant pathogens of scientific and economic importance [3]. The phytopathogen C. gloeosporioides is one of the most problematic and economically harmful phytopathogens that cause anthracnose diseases, especially in the tropic and subtropic regions of the world [4][5][6][7]. Recently, anthracnose diseases caused by C. gloeosporioides were reported to cause anthracnose diseases in fruit trees, including walnut (Juglans regia L.), and jujube (Zizyphus jujuba Miller var. ...
Colletotrichum gloeosporioides is the most prevalent phytopathogen, causing anthracnose disease that severely affects the production of various fruit trees, including walnut and jujube. In this study, the volatile organic compounds (VOCs) from Bacillus velezensis CE 100 disrupted the cell membrane integrity of C. gloeosporioides and reduced the spore germination by 36.4% and mycelial growth by 20.0% at a bacterial broth concentration of 10%, while the control group showed no antifungal effect. Based on the headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS) analysis, seven VOCs were identified from the headspace of B. velezensis CE 100. Out of the seven VOCs, 5-nonylamine and 3-methylbutanoic acid were only detected in the headspace of B. velezensis CE 100 but not in the control group. Both 5-nonylamine and 3-methylbutanoic acid showed significant antifungal activity against the spore germination and mycelial growth of C. gloeosporioides. Treatment with 100 µL/mL of 5-nonylamine and 3-methylbutanoic acid suppressed the spore germination of C. gloeosporioides by 10.9% and 30.4% and reduced mycelial growth by 14.0% and 22.6%, respectively. Therefore, 5-nonylamine and 3-methylbutanoic acid are the potential antifungal VOCs emitted by B. velezensis CE 100, and this is the first report about the antifungal activity of 5-nonylamine against C. gloeosporioides.
... The resistance to rice sheath blight Rhizocotonia solani and sheath rot Rocladittm oryzae, is improved by the overexpression of TLPs in elite indica rice cultivars (Kalpana et al., 2006). Transgenic cassava with an overexpressing rice TLP gene significantly delayed anthracnose disease and enhanced fungal tolerance compared with wild types (Odeny Ojola et al., 2018). Moreover, an overexpression of the grape TLP gene in A. thaliana resulted in its more robust resistance to powdery mildew and Pseudomonas syringae pv. ...
Hickory (Carya cathayensis) is a critical tree species of the genus Carya from the Juglandaceae family that contains nutrient-rich nuts. Due to large-scale soil degradation, the pests and diseases of hickory are becoming more and more serious. Thaumatin-like proteins (TLPs) are vital proteins involved in the complex defense process of plant pathogens. In this study, 40 CcTLP genes were identified genome-widely and phylogenetically grouped into three subfamilies. The sequence of CcTLPs had a conservative pattern, such as eight stable disulfide bonds, REDDD, and G-X-[GF]-X-C-X-T-[GA]-D-C-X(1,2)-G-X-(2,3)-C structure. In total, 57 cis-elements related to stress-responsive, light-responsive, phytohormone-responsive, and plant-responsive were discovered. Under salicylate (SA), methyl jasmonate (MeJA), and ethephon (ETH) treatments, the expressions of CcTLP28, CcTLP29, CcTLP30, CcTLP31, CcTLP32, CcTLP33, CcTLP37, CcTLP38, and CcTLP39 had different patterns. This is an indication that most of the TLP genes were upregulated by SA and downregulated by MeJA. Notably, seven TLP genes were significantly upregulated under the Botryosphaeria dothidea inoculation, especially CcTLP31, with an over 20-fold change. Nine genes were shown by subcellular localization analysis to be located at the plasma membrane and cytoplasm. The knowledge of the disease-resistant function of the CcTLP family in hickory is promoted by these results. A foundation reference for the molecular breeding of this plant in the future is provided by our findings.
... Transgenic experiments for improving agronomic traits of crops have been mostly successful in plant species for which transformation has been perfected. Cassava with enormous bioenergy potential are recalcitrant to genetic transformation as the protocols optimized are cultivar dependent (Elegba et al., 2021;Liu et al., 2011;Nyaboga et al., 2013Nyaboga et al., , 2015Ojola et al., 2018;Walsh et al., 2019). Plant beneficial endophytes offer environment-friendly methods for improving plant biomass which is advantageous for bioenergy production. ...
Cassava (Manihot esculenta. Crantz) is a starch‐rich, woody tuberous, root crop important for food, with little being done to investigate its potential as a bioenergy crop despite its enormous potential. The major bottleneck in the crop being able to serve this dual role is the competition of its storage roots for both purposes. The major cassava production regions primarily use the tuberous roots for food, and this has resulted in its neglect as a bioenergy crop. The use of non‐food cassava parts as a feedstock in cellulosic biofuel production is a promising strategy that can overcome this challenge. However, in non‐tuber parts, most of the sugars are highly sequestered in lignin complexes making them inaccessible to bacterial bioconversion. Additionally, cassava production in these major growing areas is not optimal owing to several production constraints. The challenges affecting cassava production as a food and bioenergy crop are interconnected and therefore need to be addressed together. Cassava improvement against biotic and abiotic stresses can enhance productivity and cater for the high demand of the roots for food and bioenergy production. Furthermore, increased production will enhance the usability of non‐food parts for bioenergy as the bigger goal. This review addresses efforts in cassava improvement against stresses that reduce its productivity as well as strategies that enhance biomass production, both important for food and bioenergy. Additionally, prospective strategies that could ease bioconversion of cassava for enhanced bioenergy production are explored. Cassava is very important as a food crop and has enormous potential for use in bioenergy. Majority of cassava growing regions cultivate it for food. This reviews analyses approaches that can improve cassava and make it serve a dual role of food and bioenergy.
... Constitutive expression of TLP genes enhances plant tolerance to fungal pathogens [16][17][18][19][20][21][22][23][24]. Thus, transgenic tobacco plants overexpressing peanut, rice, or cotton TLPs show increased resistance to Botrytis cinerea, R. solani, Fusarium oxysporum, F. solani [23], A. alternata [17], and Verticillium dahliae [24], whereas transgenic potatoes overexpressing the Camellia sinensis TLP gene are resistant to Phytophthora infestans [19]. ...
... Ershuizao by the absence of the CGTCA-motif and one of the six LTR-elements (Figure 8). In general, the sequences of the regulatory regions in AsTLP7, 16,20,21, and 23 were the same in cv. Sarmat and Strelets, but differed from those in cv. ...
Plant antifungal proteins include the pathogenesis-related (PR)-5 family of fungi- and other stress-responsive thaumatin-like proteins (TLPs). However, the information on the TLPs of garlic (Allium sativum L.), which is often infected with soil Fusarium fungi, is very limited. In the present study, we identified 32 TLP homologs in the A. sativum cv. Ershuizao genome, which may function in the defense against Fusarium attack. The promoters of A. sativumTLP (AsTLP) genes contained cis-acting elements associated with hormone signaling and response to various types of stress, including those caused by fungal pathogens and their elicitors. The expression of AsTLP genes in Fusarium-resistant and -susceptible garlic cultivars was differently regulated by F. proliferatum infection. Thus, in the roots the mRNA levels of AsTLP7–9 and 21 genes were increased in resistant and decreased in susceptible A. sativum cultivars, suggesting the involvement of these genes in the garlic response to F. proliferatum attack. Our results provide insights into the role of TLPs in garlic and may be useful for breeding programs to increase the resistance of Allium crops to Fusarium infections.
... In recent years, the biological roles of TLPs have become clearer. Studies have found that TLPs possess antifungal activity (Ojola et al. 2018;Sun et al. 2020), glucanase activity (Osmond et al. 2001; Menu-Bouaouiche (Palacín et al. 2010;Kumar et al. 2013). TLPs are also involved in many facets of both growth and development. ...
Main conclusion
A total of 27 rose thaumatin-like protein (TLP) genes were identified from the rose genome through bioinformatics analyses. RcTLP6 was found to confer salinity stress tolerance in rose.
Abstract
Thaumatin-like proteins (TLPs) play critical roles in regulating many biological processes, including abiotic and biotic stress responses in plants. Here, we conducted a genome-wide screen of TLPs in rose (Rosa chinensis) and identified 27 RcTLPs. The identified RcTLPs, as well as other TLPs from six different plant species, were placed into nine groups based on a phylogenetic analysis. An analysis of the intron–exon structures of the TLPs revealed a high degree of similarity. RcTLP genes were found on all chromosomes except for chromosome four. Cis-regulatory elements (CEs) were identified in the promoters of all RcTLPs, including CEs associated with growth, development and hormone-responsiveness, as well as abiotic and biotic responses, indicating they play diverse roles in rose. Transcriptomics analysis revealed that RcTLPs had tissue-specific expression patterns, and several root-preferential RcTLPs were responsive to drought and salinity stress. Quantitative PCR analysis of six RcTLPs under ABA, PEG and NaCl treatment confirmed the differentially expressed genes identified in the transcriptomics experiment. In addition, silencing RcTLP6 in rose leaves led to decreased tolerance to salinity stress. We also screened proteins which may interact with RcTLP6 to understand its biological roles. This study represents the first report of the TLP gene family in rose and expands the current understanding of the role that RcTLP6 plays in salt tolerance. These findings lay a foundation for future utilization of RcTLPs to improve rose abiotic stress tolerance.
