Use of plant extracts and biocontrol agents for the management of brown spot disease in rice

Fifty plant extracts, four oil cakes and eight antagonistic organisms were tested against Bipolaris oryzae (Cochliobolus miyabeanus), the causal agent of brown spot disease of rice. In vitro studies indicated that two leaf extracts, Nerium oleander and Pithecolobium dulce exerted the higher percent inhibition to mycelial growth (77.4, 75.1%) and spore germination (80.3, 80.0%) of B. oryzae. Among the four oil cake extracts tested in vitro against B. oryzae, neem cake extract showed the maximum inhibition percent to mycelial growth (80.18%) and spore germination (81.13%) of the pathogen followed by mahua cake extract, castor and gingelly cake extract. Trichoderma viride (Tv2) was significantly effective in inhibiting the mycelial growth (62.92%) and spore germination (77.03%) of the pathogen followed by Trichoderma harzianum (Th5) and Trichoderma reesei (Tr3). The promising leaf extracts, oil cake extracts and antagonistic microorganisms were further evaluated for their efficacies in disease management under glasshouse and field conditions. In glasshouse studies, post-infectional spraying of rice plants with neem cake extract, N. oleander leaf extract and T. viride (Tv2) was significantly effective in reducing the incidence of brown spot of rice by 66, 52 and 45 percent respectively. Two rounds of spraying of rice plants with neem cake extract, N. oleander leaf extract and T. viride (Tv2) in the field at initial appearance of disease and 15 days later reduced the incidence of brown spot (70, 53 and 48% disease reduction respectively) and increased the yield by 23, 18 and 15 percent respectively.     

Control of storage rot of cassava tuber caused by Rhizopus oryzae using some plant extracts

Abstract

The potency of Piper nigrum seed and leaf, Aframomum meleguata seed and Ageratum conyzoides leaf extracts in the control of cassava tuber rot caused by Rhizopus oryzae was investigated. Water, ethanol and petroleum ether were used as extracting solvents. These extracts were fungitoxic both in vitro and in vivo against the test pathogen. P. nigrum seed extracts were the best followed by those of A. meleguata seed, A. conyzoides leaf and then P. nigrum leaf. The extracts were more effective in controlling rot development in unwounded than wounded tubers especially when they were applied before inoculation with the test pathogen. Ethanol extracts gave the highest growth inhibition in vivo followed by water and the petroleum ether extract whereas water extracts showed marked superiority over the extracting solvents in checking rot development in cassava tuber. Water and ethanol extracts of P. nigrum seed and leaf, A. meleguata seed and A. conyzoides leaf could be used as pesticide of plant origin in the control of R. oryzae causing cassava tuber rot in storage.     

Fungicidal activity of some plant extracts against rhizoctonia

The leaf extracts of Piper guinensis, Ocimum sanctum, O. gratissimum and Cymbopogon citratus were effective in reducing the radial growth of Rhizoctonia solani in vitro and in checking the spread of the disease caused by the pathogen in the field. P. guinensis was the best in checking the spread of the pathogen both in vitro and in vivo. This was followed by C. citratus, O. sanctum and O. gratissimum. Citrus limon reduced the radial growth of the pathogen in vitro but failed to check the spread of the disease effectively in the field. Alcohol extracts were more effective than water extracts.     

Antifungal potential of some higher plants against Fusarium udum causing wilt disease of Cajanus cajan

The fungitoxic effects of different plant extracts on Fusarium udum, which causes wilt disease of Cajanus cajan in vitro and in vivo, were examined. The complete arrest of the radial growth of the pathogen occurred at a 10% concentration of leaf extract from Adenocallyma alliaceum. A leaf extract of Citrus medica, a root extract of Asparagus adscendens, rhizome extracts of Curcuma longa and Zingiber officinale, and a bulb extract of Allium sativum inhibited up to 100% growth at higher concentrations. A. alliaceum controlled the disease up to 100% by amending its 4% powder in unsterilized soil and 2% in sterilized soil. The population of F. udum was found to be markedly reduced following treatments with plant powders.     

A comparative screening of hybrid,modern varieties and local rice cultivar for brown leaf spot disease susceptibility and yield performance

Five rice cultivars, one hybrid (WR96), three modern (BR16, BR26, and BRRI Dhan27) and one local (Pari) were screened for reaction to brown leaf spot disease caused by Cochliobolus miyabeanus and performance of yield-related characters. The severity of brown leaf spot varied with growth stages of rice plant as well as different cultivars tested under field condition. Low disease severity was observed at maximum tillering stage compared to moderate to high at dough stage, with hybrid cultivar WR96 showing highest disease, while local cultivar Pari had the lowest. Brown spot disease severity in different cultivars under induced epiphytic condition also followed the similar trend. The results also revealed that most of the yield-contributing characters examined showed wide variations among the cultivars. Modern cultivar BR16 produced the highest panicle length, number of grain per panicle and grain yield per hectare. At the same time as local cultivar Pari generated the lowest number of tiller per plant, panicle length, grain number per panicle and grain yield per hectare. Moreover, hybrid cultivar WR96 produced the highest percentage of spotted grain per panicle and seed yielding C. miyabeanus, and also the lower percentage of seed germination, while the reverse was observed in local cultivar Pari. These findings may allow producers and breeders to select rice cultivar, resistant or tolerant to brown leaf spot disease and to avoid significant reductions in grain yields.     

Antifungal activity of extracts from five Egyptian wild medicinal plants against late blight disease of tomato

This study was carried out to evaluate the antifungal potential of water and ethanol extracts from aerial parts of five wild medicinal plants collected from Sinai Peninsula, Egypt, and the extracts were tested in vitro and in vivo against Phytophthora infestans, the causal agent of late blight disease of tomato. The five wild medicinal plants used for the study were Asclepias sinaica, Farsetia aegyptia, Hypericum sinaicum, Phagnalon sinaicum and Salvia aegyptiaca. Ethanol extracts were more effective on the pathogen than water extracts at all concentrations used. Water and ethanol extracts of all plants tested reduced mycelial growth and inhibited spore germination of the pathogen with varying degrees. Water and ethanolic extracts also reduced the disease infection with pathogen comparing with control in detached leaves technique. In all experiments, extracts of A. sinaica and F. aegyptia were most effective on the pathogen than other plant extracts. In plot experiment, the water and ethanolic extracts of A. sinaica and F. aegyptia gave the most reduction of late blight disease severity comparing with control. Also, data indicated that after the application of plant extracts, there was an increase in fruit yield of tomato corresponding with the reduction of disease severity . Scanning electron microscopy revealed a negative alteration of pathogen hyphae treated with A. sinaica extract at 20%. This concentration of the same plant extract also reflected in dramatic changes in the cyto-morphology of pathogen hyphae as observed by transmission electron microscopy. These changes resulted in an increase in vacuolisation and lipid contents with consequent reduction of cytoplasm with alteration of cell wall and plasmalemma. The overall results suggested that the use of these Egyptian wild medicinal plant extracts was promising, effective and environment-friendly management measure against Phytophthora blight of tomato and thus, may be used in the production of organically grown vegetables.     

Fungitoxic effects of some leaf extracts against rhizopus oryzae causing tuber rot of potato

Extracts of Cymbopogon citratus, Azadirachta indica (Neem) and Ocimum gratissimum were effective in controlling the growth of Rhizopus oryzae in vitro and in vivo. Neem was found to be the best followed by C. citratus and O. gratissimum. Alcohol extracts of these plant species were more effective than water extracts in reducing the growth of the pathogen in culture and in checking rot development in potato tuber by the fungus. A. indica and C. citratus have considerable disease control potential and their extracts can be exploited as a source of pesticide of plant origin to control the soft rot in stored potato incited by Rhizopus oryzae.     

Inhibitory effect of botanical extracts against Alternaria alternata of aloe vera dry rot

Aloe vera dry rot caused by Alternaria alternata, is one of the most serious fungal diseases affecting the commercial cultivation of aloe vera in North India. Control of this disease through chemical is quite expensive and not ecofriendly. The plant extracts as biopesticdes act as a vital components for the management of this disease. Evaluation of some plant extracts was carried out against A. alternata in laboratory conditions. The extracts were prepared at 5% and 10% concentration and were evaluated through inhibition in radial growth (food poison technique) and spores (conidia) germination (hanging drop technique) against A. alternata. Neem leaf extract gave 58.6% inhibition in radial growth and 56.5% in spore germination at 10% concentration followed by Ocimum sanctum which was found effective and gave 54.7% inhibition in radial growth and 50.4% in spore germination over control.     

Effect of Piper betle L. and its extracts on the growth and aflatoxin production by Aspergillus parasiticus

Ground powder of the leaf and fruit of Piper betle L., a tropical spice plant grown in Southeast Asia, was prepared and extracted by chloroform, ethanol and water with one solvent only or with 3 solvents in sequence. The betel powder and various extracts were added to YES broth to determine their effects on the growth and aflatoxin production by Aspergillus parasiticus. Results showed that betel leaf powder exhibited higher antimycotic activity than fruit. One half percent of ground leaf powder completely inhibited the growth and aflatoxin production by A. parasiticus. Among the solvent extracts, chloroform and ethanol extracts of betel leaf prepared from a single solvent extraction showed more antimycotic activity. The ethanol extract of betel leaf at the level of 450 micrograms/ml would eliminate A. parasiticus growth and aflatoxin production. The antimycotic activity of this ethanol extract was most pronounced at pH 4.     

Soil moisture mediated interaction between <Emphasis Type="Italic">Polygonatum biflorum</Emphasis> and leaf spot disease

Fungal pathogens can regulate the abundance and distribution of natural plant populations by inhibiting the growth, survival, and reproduction of their hosts. The abiotic environment is a crucial component in host–pathogen interactions in natural plant populations as favorable conditions drive pathogen development, reproduction, and persistence. Foliar plant pathogens, such as fungal lesions referred to generically as “leaf spot disease,” are particularly responsive to increased moisture levels, but the manner in which the abiotic environment drives disease dynamics, and how these diseases regulate natural plant populations, is not fully understood. We investigate (1) the impact of ambient soil moisture and diffuse light on the prevalence of a leaf spot pathogen (Phyllosticta sp.) in a natural population of Polygonatum biflorum, an understory herb native to deciduous forest understories in the eastern US, and (2) the effects of the fungal pathogen on the survival, growth, and abundance of the plants. We tracked six P. biflorum populations and disease incidence, as well as soil moisture and diffuse light, between 2003 and 2005 in the understory deciduous forest of the southern Appalachian Mountains, North Carolina, USA. Results show that both the occurrence of P. biflorum and the prevalence of P. biflorum leaf spot disease are highest where soil moisture is intermediate and diffuse light is lowest. Disease occurrence depends upon plant presence, but it also adversely impacts plant survival, abundance, and growth. These results suggest that leaf spot disease is likely to impact population dynamics, which in turn vary as a function of environmental drivers.     

Effects of Slag-Based Silicon Fertilizer on Rice Growth and Brown-Spot Resistance

It is well documented that slag-based silicon fertilizers have beneficial effects on the growth and disease resistance of rice. However, their effects vary greatly with sources of slag and are closely related to availability of silicon (Si) in these materials. To date, few researches have been done to compare the differences in plant performance and disease resistance between different slag-based silicon fertilizers applied at the same rate of plant-available Si. In the present study both steel and iron slags were chosen to investigate their effects on rice growth and disease resistance under greenhouse conditions. Both scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to examine the effects of slags on ultrastructural changes in leaves of rice naturally infected by Bipolaris oryaze, the causal agent of brown spot. The results showed that both slag-based Si fertilizers tested significantly increased rice growth and yield, but decreased brown spot incidence, with steel slag showing a stronger effect than iron slag. The results of SEM analysis showed that application of slags led to more pronounced cell silicification in rice leaves, more silica cells, and more pronounced and larger papilla as well. The results of TEM analysis showed that mesophyll cells of slag-untreated rice leaf were disorganized, with colonization of the fungus (Bipolaris oryzae), including chloroplast degradation and cell wall alterations. The application of slag maintained mesophyll cells relatively intact and increased the thickness of silicon layer. It can be concluded that applying slag-based fertilizer to Si-deficient paddy soil is necessary for improving both rice productivity and brown spot resistance. The immobile silicon deposited in host cell walls and papillae sites is the first physical barrier for fungal penetration, while the soluble Si in the cytoplasm enhances physiological or induced resistance to fungal colonization.     

Antibacterial activity of certain plant extracts against bacterial wilt of tomato

Abstract

Five isolates of Ralstonia solanacearum were isolated from a naturally wilted root of tomato plants grown in Assiut governorate. The antibacterial activity of extract of Datura, Garlic and Nerium were tested in controlling R. solanacearum in vitro and in vivo. Garlic exhibited the strongest antibacterial activity against bacterial wilt in vitro and in vivo followed by Datura and then Nerium. Cold water extracts of these plant species were more effective than hot water extract in the development of the disease in vivo. In greenhouse experiments, the application of the tested plant extract to soil at the time of inoculation, two days before inoculation and two days after inoculation the pathogen, significantly reduced the disease index of wilt on Super Marmande tomato cultivars. The application of plant extracts at the same time as inoculation resulted in the highest reduction of disease index.     

Allelopathic effect of Nymphaea lotus L. on growth and yield of cultivated rice around Lake Manzala (Nile Delta)

Lotus infestation of ricefields is a major cause of crop failure and decrease of grain yield in the newly reclaimed cut-off areas around lake Manzala, Egypt. This study provides insights in the allelopathic effect of Nymphaea lotus L. on rice (Oryza sativa cavr. Giza-177). Lotus rhizome extracts were inhibitory to seed germination and seedling growth of rice. The degree of inhibition was dependent on extract type and concentration. Ethanol and water extracts were more inhibitory than chloroform extracts. The phenolic fraction of ethanol extracts showed the highest inhibitory effects. In a target (rice)-neighbour (lotus) pot experiment, rice dry mass and relative growth rate were dependent on its age and on lotus rhizome density, with decreased growth at increased lotus density. Field data on infested and non-infested ricefields demonstrated a decreased leaf area index and yield in infested fields. Identification of the potential allelochemical compounds by gas chromatography/mass spectrometry revealed the presence of allelopathic phenolics in lotus rhizomes.     

泽泻汤提取物三萜类化学成分分析及其抗非酒精性脂肪肝作用研究

目的:对泽泻汤不同极性溶剂提取物三萜类化学成分进行定性分析并探究其对非酒精性脂肪肝的干预作用。方法:制备泽泻汤水及95%乙醇提取物,利用超高效液相色谱串联高分辨质谱技术对三萜类成分进行定性鉴别,并比较两种提取物中三萜类化合物数量及含量差异。采用油酸诱导的HepG2细胞脂肪累积模型评价泽泻汤不同极性提取物干预非酒精性脂肪肝的能力。结果:在泽泻汤中共鉴定20个三萜类色谱峰,水提取物中鉴定出14个三萜类成分,其中16-oxo-alisol A相对含量最高,95%乙醇提取物中鉴定出16个三萜类成分,alisol B 23-acetate相对含量最高。两提取物均能显著降低细胞中的脂肪累积量,且乙醇提取物表现出更强的抑制细胞内脂肪累积的作用。结论:泽泻汤水及95%乙醇提取物中三萜类化合物数量和含量存在差异,两种提取物均具有潜在抗非酒精性脂肪肝作用,且乙醇提取物活性更强,研究结果为泽泻汤进一步的药效物质基础及药理活性研究提供了基础。     

Cloning and overexpression of antifungal barley chitinase gene in Escherichia coli

Plant chitinases are pathogenesis-related proteins, which are believed to be involved in plant defense responses to pathogen infection. In this study, chitinase gene from barley was cloned and overexpressed in Escherichia coli. Chitinase (35 kDa) was isolated and purified. Since the protein was produced as insoluble inclusion bodies, the protein was solubilized and refolded. Purified chitinase exerted broad-spectrum antifungal activity against Botrytis cinerea (blight of tobacco), Pestalotia theae (leaf spot of tea), Bipolaris oryzae (brown spot of rice), Alternaria sp. (grain discoloration of rice), Curvularia lunata (leaf spot of clover) and Rhizoctonia solani (sheath blight of rice). Due to the potential of broad-spectrum antifungal activity barley chitinase gene can be used to enhance fungal-resistance in crop plants such as rice, tobacco, tea and clover.     

Integration of plant growth-promoting rhizobacteria and chemical elicitors for induction of systemic resistance in mulberry against multiple diseases

Abstract

In mulberry (Morus alba L.), various individual strains of plant growth-promoting rhizobacteria (PGPR) and synthetic analogs of naturally occurring plant activators have demonstrated their potential to elicit induced systemic resistance (ISR) against either brown leaf spot (Cercospora moricola) or leaf rust (Cerotelium fici) diseases. However, these biological and chemical elicitors have not been evaluated so far against multiple infections of both these diseases which commonly occur during the post-rainy season. The present study was therefore aimed to assess the capability of PGPR strains and chemical plant activators, as individual and in integration, in elicitation of ISR against multiple infections. Three PGPR strains, Azotobacter chroococcum strain Azc-3, Bacillus megaterium strain Bm-1 and Pseudomonas fluorescens strain Psf-4, and plant activators, acetyl-salicylic acid (ASA), sodium salicylate (NaS) and 4-amino-n-butyric acid (ABA) were selected for the study. Under in vitro tests, all the plant activators up to 2000 ppm concentration exhibited their compatibility with the PGPR strains tested. Upon assaying of elicitors with plant-pathosystem, disease suppression was significantly (p = 0.05) high with integrated application of PGPR strains and plant activators when compared to their individual applications. All the elicitors at individual application varied in their response to multiple infections with the plant age. However, integration of Azc-3 + ASA provided greater suppression to multiple infections of brown leaf spot and leaf rust diseases during the entire growth period of mulberry plants. Thus, this combination of biological and chemical elicitors holds great promise to provide an effective ecofriendly alternative to the toxic chemical fungicides presently recommended for the control of brown leaf spot and leaf rust diseases in mulberry.     

Cocculus hirsutus extract inhibits the Xanthomonas oryzae pv. oryzae,the bacterial leaf blight pathogen in rice

Plant-derived natural bactericides and their possible applications in agriculture to control plant bacterial diseases has intensified as this approach has enormous potential to inspire and influence modern agro-chemical research. Naturally occurring and biologically active plant products such as essential oils and organic extracts could be a source of alternative classes of natural biopesticides to serve as templates for new and more effective compounds in controlling plant pathogenic micro-organisms. In the present study, the efficacy of six plants extracts from different solvent system were tested for their antibacterial activity aganist Xanthomonas oryzae pv. oryzae both in vitro and in vivo. Among these extracts, Cocculus hirsutus leaf chloroform extract exhibits significant antibacterial activity against X. oryzae pv. oryzae. Data obtained from the experiments such as minimum inhibitory concentration, effect of C. hirsutus leaf chloroform extract on the incidence of X. oryzae pv. oryzae, phytotoxicity test and effect of C. hirsutus leaf chloroform extract on seed germination and seedling vigour, along with the in vivo experiments under greenhouse conditions showed significant improvement over controls. Thus, the present study demonstrated that the C. hirsutus leaf chloroform extract posses antibacterial activity against bacterial leaf blight pathogen of rice.     

Rice Resistance to Brown Spot Mediated by Silicon and its Interaction with Manganese

Brown spot is one of the most devasting and prevalent disease of rice and its control is mainly dependent on fungicide application. Therefore, this study aimed to examine the effects of Si and Mn on the development of brown spot on rice plants grown in hydroponic culture. Rice plants (cv. ‘Metica‐1’) were grown in plastic pots containing 0 or 2 mm Si (?Si and +Si treatments, respectively) with three Mn rates (0.5, 2.5 and 10 μm ). Plants were inoculated with B. oryzae 39 days after emergence. The following components of resistance were evaluated: incubation period (IP), number of lesions (NL) per cm² of leaf area, real disease severity (RDS) and area under brown spot progress curve (AUBSPC). The content of Si and Mn in the plant tissues was also determined. Si content was significantly higher in rice tissue of plants of the +Si treatment than of the ?Si treatment regardless of the Mn rates used. The Mn rates did not affect the Si content of the rice plants. The Mn content of the rice tissues was significantly higher in the ?Si treatment than on +Si treatment, regardless of the Mn rate used. The Mn content was significantly lower at 0.5 μm Mn in comparison to the other rates for both ?Si and +Si treatments. The IP of brown spot on rice leaves significantly increased in the +Si treatment; but the Mn rates in the presence of Si had no effect on IP. In the ?Si treatment, the IP was significantly higher only at the rate of 2.5 μm . The NL, RDS and AUBSPC were significantly reduced in the +Si treatment regardless of the Mn rates. The Mn rates in the presence of Si had no effect on these components of resistance. Overall, Si dramatically impacted the development of brown spot regardless of the presence of Mn at either low or high rates. This may be useful in regions where the soil has either toxic or deficient levels of Mn and cultivars with brown spot resistance are not commercially available.     

Effect of Zinc on the Development of Brown Spot in Rice

Brown spot, caused by the fungus Bipolaris oryzae, is one of the most destructive diseases of rice. This study investigated the effect of zinc rates on the development of brown spot in rice. Rice plants (cv. ‘Metica‐1′) were grown in hydroponic culture amended with Zn rates (applied as ZnSO₄^.7H₂O) of 0, 0.5, 1, 2 and 4 μm and inoculated with B. oryzae. The foliar concentration of Zn was determined. Leaf samples were assessed for disease severity, and then, area under brown spot progress curve (AUBSPC) was calculated. The relationship between Zn concentrations on leaf tissues and the rates of this micronutrient was best described by a positive linear regression model, while the relationship between the Zn rates and the AUBSPC was best described with a positive quadratic regression model. The correlation between Zn concentrations on leaf tissues and AUBSPC was positive and significant (r = 0.68, P < 0.05). The results from this study showed that high foliar concentration of Zn was associated with increasing rice susceptibility to brown spot.