Toxicological effects of nanoparticles in plants: Mechanisms involved at morphological, physiological, biochemical and molecular levels.

The rapid advancement of nanotechnology has led to unprecedented innovations across diverse industries, including pharmaceuticals, agriculture, cosmetics, electronics, textiles, and food, owing to the unique properties of nanoparticles. The extensive production and unregulated release of synthetic nanoparticles may contribute to nanopollution within the ecosystem. In the agricultural sector, nanotechnology is increasingly utilized to improve plant productivity, enhance resistance to stressors, and reduce the usage of chemicals. However, the uncontrolled discharge of nanoparticles into the natural environment raises concerns regarding possible plant toxicological impacts. The review focuses on the translocation of these particles within the plants, emphasizing their phytotoxicological effects at morphological, physiological, biochemical, and molecular levels. Eventhough the beneficial aspects of these nanoparticles are evident, excessive usage of nanoparticles at higher concentrations may lead to potential adverse effects. The phytotoxicity resulting from excessive amounts of nanoparticles affects seed germination and biomass production, disrupts the photosynthesis system, induces oxidative stress, impacts cell membrane integrity, alters gene expression, causes DNA damage, and leads to epigenetic variations in plants. Nanoparticles are found to directly associate with the cell membrane and cell organelles, leading to the dissolution and release of toxic ions, generation of reactive oxygen species (ROS) and subsequent oxidative stress. The present study signifies and accumulates knowledge regarding the application of nanoparticles in agriculture and illustrates a clear picture of their possible impacts on plants and soil microbes, thereby paving the way for future developments in nano-agrotechnology. The review concludes by addressing current challenges and proposing future directions to comprehend and mitigate the possible biological risks associated with nanoparticles in agriculture.

Fate, bioaccumulation and toxicity of engineered nanomaterials in plants: Current challenges and future prospects.

The main focus of this review is to discuss the current advancement in nano-metallic caused phytotoxicity on living organisms and current challenges in crops. Nanostructured materials provide new tools in agriculture to boost sustainable food production, but the main concern is that large-scale production and release of nanomaterials (NMs) into the ecosystem is a rising threat to the surrounding environment that is an urgent challenge to be addressed. The usage of NMs directly influences the transport pathways within plants, which directly relates to their stimulatory/ inhibitory effects. Because of the unregulated nanoparticles (NMs) exposure to soil, they are adsorbed at the root surface, followed by uptake and inter/intracellular mobility within the plant tissue, while the aerial exposure is taken up by foliage, mostly through cuticles, hydathodes, stigma, stomata, and trichomes, but the actual mode of NMs absorption into plants is still unclear. NMs-plant interactions may have stimulatory or inhibitory effects throughout their life cycle depending on their composition, size, concentration, and plant species. Although many publications on NMs interactions with plants have been reported, the knowledge on their uptake, translocation, and bioaccumulation is still a question to be addressed by the scientific community. One of the critical aspects that must be discovered and understood is detecting NMs in soil and the uptake mechanism in plants. Therefore, the nanopollution in plants has yet to be completely understood regarding its impact on plant health, making it yet another artificial environmental influence of unknown long-term consequences. The present review summarizes the uptake, translocation, and bioaccumulation of NMs in plants, focusing on their inhibitory effects and mechanisms involved within plants.

Induction of drought tolerance in Pennisetum glaucum by ACC deaminase producing PGPR- Bacillus amyloliquefaciens through Antioxidant defense system.

Rhizobacteria from pearl millet were screened to produce 1-aminocyclopropane-1-carboxylate (ACC) deaminase and to evaluate its role in alleviating drought stress. Amongst 96 isolates, 28 were positive for ACC deaminase production, with MMR04 offering maximum activity of 2196.23 nmol of α-ketobutyrate produced mg-1 of protein h-1. The ACC deaminase producing rhizobacteria with multiple beneficial properties along with root colonization and non-pathogenic were selected [Bacillus amyloliquefaciens (MMR04), Bacillus subtilis (MMR18) and Stenotrophomonas maltophilia (MMR36)] to confirm the presence of ACC deaminase gene. A significant enhancement in seed germination (91.75%) and seedling vigor (1213.73) was noted upon seed treatment with MMR04 and hence further evaluated for its ability to induce drought stress. The seed treatment with MMR04 improved plant growth parameters and total chlorophyll and RWC in plants grown under severe drought stress (G5) conditions compared to control plants. In addition, MMR04 seed treatment enhanced proline, APX and SOD activity while decreased the MDA content up to 2.3 fold compared to untreated plants (G5). Gene expression studies revealed a significant decrease of 3.3 and 1.8 fold in the relative expression of drought-responsive (DREB-1E) and ethylene-responsive factor (ERF-1B) marker genes, respectively and an increase of 2.2 and 2.9 fold in the relative expression of APX1 and SOD1, respectively in MMR04 treated plants grown under G5 conditions over control. The results confirmed that ACC deaminase producing B. amyloliquefaciens MMR04 could defend the pearl millet plants against drought stress through an antioxidative system, thereby warranting its application in drought stress management.

Morphological and molecular characterization of Neopestalotiopsis vitis associated with leaf blight disease of Manilkara zapota-a new record from India.

Sapota is an important horticultural crop grown in India, and Karnataka is a major producer of sapota. A characteristic leaf blight disease was observed in Southern Karnataka during field surveys conducted in 2019 with an incidence of 13-22% in approximately 45 ha of sapota field. The leaf blight-associated pathogen was isolated on the potato dextrose agar medium. A total of 12 isolates obtained from each location were identified culturally and morphologically. Based on the morphological and cultural features, the pathogen was identified as Pestalotiopsis or Neopestalotiopsis, which was further confirmed by molecular identification using a representative isolate (MZ03). The ITS rDNA and ß-tubulin genes were amplified and sequenced using ITS1/ITS4 and T1/T22 primer pairs respectively. nBLAST search analysis and concatenated (ITS-rDNA and TUB2 loci) phylogenetic analysis confirmed the pathogen identity as Neopestalotiopsis vitis. Pathogenicity tests conducted on detached leaves by inoculation with a conidial suspension of N. vitis produced typical blight symptoms after 4-5 days and progressed to cover the entire leaf lamina after 10-12 days. The pathogen's identity was confirmed after re-isolation by cultural and morphological features. Although Pestalotiopsis clavispora and Pestalotiopsis versicolor causing diseases on sapota seedlings and trees have been reported, no reports are available for the occurrence of N. vitis to sapota from India. This is the first report of N. vitis associated with leaf blight disease of sapota from India.

Dactuliophora mysorensis sp. nov.: A New Species of Mycelia Sterilia Causing Zonate Leaf Spot on Cowpea in India.

Cowpea is an important pulse crop extensively grown in arid and semi-arid tropics which is affected by a number of diseases. Fungi belonging to mycelia sterilia are known to cause many diseases on cereals and pulses. During the cowpea field survey in Mysore District of Karnataka (India), Dactuliophora sp. was identified as the major pathogen causing zonate leaf spot (ZLS) disease. The fungal pathogen was isolated from naturally infected cowpea leaves and identified as a member belongs to the genus Dactuliophora, which was previously described by CLA Leakey in the year 1964 on Vigna unguiculata from Africa. However, detailed morphological and cultural examinations of the pathogen revealed striking differences from that of D. tarrii. Based on differences in morphology with D. tarrii, a new species Dactuliophora mysorensis sp. nov. is described herein. The disease incidence as well as disease index was estimated for 3 years (2016-2018). The severity of the disease was high during August-November. High incidence and disease index of ZLS was recorded in Doddamaragowdanahally region. The pathogenicity tests demonstrated similar symptoms of ZLS. The ITS barcoding revealed that the pathogen is closely related to Rhizoctonia bataticola and Macrophomina phaseolina. Further, in vitro evaluation of fungicides was carried out by poisoned food technique. Among the five fungicides examined, only two systemic fungicides (Benomyl and Carbendazim) were effective against D. mysorensis. Thus, the present study recommends Benomyl and Carbendazim for management of ZLS disease caused by D. mysorensis.

First Report of Nigrospora sphaerica associated with Leaf Spot Disease of Cowpea (Vigna unguiculata) from India.

Cowpea (Vigna unguiculata (L.) Walp) is one of the main legume crops grown in arid and semi-arid regions in the world. Brazil, Haiti, Myanmar, Nigeria, Sri Lanka, United States, and India contributes to the substantial production of cowpea at the global level (Mahadevakumar and Janardhana, 2012, 2014). Field surveys conducted during 2017-19 (August-September) in major cowpea growing regions of southern Karnataka revealed the occurrence of characteristic leaf spot disease of unknown etiology with an incidence ranging from 6 to 8%. Initially, the symptoms developed as small specks (1.5 to 3.5 mm), characterized by circular or irregular shape. These lesions began to develop from the leaf margin and regularly extended and coalesced to form larger lesions. After the successful manifestation of the symptoms on leaves, the associated fungal pathogen was isolated. In brief, the infected leaves were surface sterilized with 2% NaOCl for 2 min, rinsed thrice in sterile distilled water (SDW) and blotter dried. The leaf sections were placed on potato dextrose agar (PDA) in Petri plates and incubated at room temperature (27 ± 2°C) for 10 to 12 days. Mycelia developed from infected tissues were transferred to fresh PDA plates and pure cultures were obtained. Mycelia were initially white and eventually turned into gray. The conidia were black, single-celled, smooth, spherical to subspherical, 10 to 22 µm in diameter (n=30), and borne singly on a hyaline vesicle at the tip of each conidiophore. Based on the cultural features and conidial morphology, the fungus was identified as Nigrospora sp. Further, to identify the pathogen to the species level, the ITS region of the ribosomal RNA gene was amplified using primers ITS1 and ITS4 (White et al. 1990). The amplified PCR products were purified and sequenced. The nBLAST analysis showed 100% similarity with reference sequences from the GenBank database Nigrospora sphaerica (MT225783.1; MN795578.1), and the sequences were deposited in GenBank (Accession No. MT305812.1, MT305813.1, MT305814.1). Based on the cultural, morphological, microscopic and molecular characteristics, the associated fungal pathogen was identified as N. sphaerica (Sacc.) Mason (Chen et al. 2018; Wang et al. 2017) and a voucher specimen was deposited at University of Mysore Herbarium with accession No. UOM20-NS1. Further, pathogenicity tests were conducted on healthy cowpea plants grown under greenhouse conditions. Inoculations were made with conidial suspension (105 conidia/ml) prepared in SDW and healthy plants sprayed with SDW served as a standard control. All the plants were covered with polyethylene bags for 24-48 hr and observations were made at regular intervals. Typical necrotic lesions developed after 12 days of inoculation and no such symptoms were observed on the standard control set. The associated pathogen was re-isolated from diseased leaves and its identity confirmed based on morphology and cultural characteristics. Leaf spots are becoming a major problem in cowpea growing areas in recent years (Dactuliophora sp., Pestalotiopsis leaf spot, Alternaria leaf spot, and many others) (Mahadevakumar and Janardhana 2012, 2014). Recently, Aplosporella hesperidica causing collar rot on cowpea has been reported from the same region (Deepika et al. 2020). The seed borne occurrence N. sphaerica on cowpea is reported from Brazil (Rodrigues and Menezes 2002), there are no previous reports available on the occurrence of N. sphaerica on cowpea leaf spots, the present investigation is the first report of N. sphaerica causing leaf spot disease on cowpea from India.

A new collar rot disease of cowpea (Vigna unguiculata) caused by Aplosporella hesperidica in India.

Cowpea is an important pulse crop cultivated in arid and semi-arid regions of the world. During field survey, a characteristic wilt was observed in around 45 ha of cowpea fields with incidence 17-25%. Infection was seen in pre-flowering stage and infected plants showed quick wilt symptoms with tan lesions near the stem-soil interface. Fungal pathogens associated were isolated on PDA, which produced dark to grey olivaceous colonies in the centre, and aerial mycelia were appressed with floccose and white to smoke-grey. Conidia are aseptate, initially hyaline, smooth-walled, broadly ellipsoidal with rounded ends becoming dark brown. Based on these morphological features, the fungal pathogen was identified as Aplosporella sp. The ITS-rDNA region was amplified using ITS1/ITS4 primers and sequenced. The nBLAST and phylogenetic analysis confirmed the pathogen as Aplosporella hesperidica. The Koch's postulates were performed on 45-days-old cowpea plants with mycelial disc of A. hesperidica. Development of typical necrotic lesions was observed after 28 days of post-inoculation and the pathogen's identity was confirmed based on re-isolation. Efficacy of fungicides evaluated in vitro showed that the pathogen is highly sensitive to systemic fungicides rather than the contact fungicides. The cowpea production was severely affected owing to the causative agent A. hesperidica. The collar rot disease of cowpea by A. hesperidica is the first report in India. SIGNIFICANCE AND IMPACT OF THE STUDY: A new collar rot disease of cowpea recorded from India has been investigated. The necrotic lesions were enlarged and eventually quick wilt and death of the host plant was observed with incidence ranged from 17 to 25%. Associated fungal pathogen was isolated and identified as Aplosporella hesperidica based on morphology and ITS-rDNA sequence analysis. Koch's postulates were performed under greenhouse conditions and in vitro evaluation of fungicides shows that the pathogen is sensitive to systemic fungicides. This is the first report of A. hesperidica causing collar rot disease of cowpea in India.

Antibacterial and antimitotic potential of bio-fabricated zinc oxide nanoparticles of Cochlospermum religiosum (L.).

Zinc oxide nanoparticles synthesized through eco-friendly approach has gained importance among researchers due to its broad applications. In the present work, hexagonal wurtzite shape nanoparticles (below 100 nm size) were obtained using aqueous leaf extract of Cochlospermum religiosum which was confirmed through X-Ray diffraction (XRD) analysis. The synthesized ZnO-NPs showed an absorption peak at 305 nm which is one of the characteristic features of ZnO-NPs.The bio-fabricated ZnO-NPs were of high purity with an average size of ∼76 nm analyzed through Dynamic Light Scattering (DLS) analysis supporting the findings of XRD. The SEM images confirmed the same with agglomeration of smaller nanoparticles. The composition of aqueous leaf extract and ZnO-NPs was explored with Fourier Transform Infrared Spectroscopy (FT-IR). The plant extract as well as bio-fabricated ZnO-NPs offered significant inhibition against Gram-positive (B. subtilis and Staph. aureus) and Gram-negative (P. aeruginosa and E. coli) bacteria. The minimum inhibitory concentration (MIC) of bio-fabricated ZnO-NPs and plant extract was found between 4.8 and 625 µg/ml against test pathogens, which was authenticated with live and dead cell analysis. Apart from antibacterial potentiality, antimitotic activity was also observed with a mitotic index of 75.42% (ID50 0.40 µg mL-1) and 61.41% (ID50 0.58 µg mL-1) in ZnO-NPs and plant extract, respectively. The results affirm that plant extract and its mediated ZnO-NPs possess biological properties.

Molecular profiling and bioactive potential of an endophytic fungus Aspergillus sulphureus isolated from Sida acuta: a medicinal plant.

CONTEXT: Sida acuta Burm.f. (Malvaceae) extracts are reported to have applications against malaria, diuretic, antipyretic, nervous and urinary diseases. No fungal endophytes of S. acuta are reported. OBJECTIVE: Isolation, identification and evaluation of antibacterial, antioxidant, anticancer and haemolytic potential of fungal endophytes from the ethnomedcinal plant S. acuta. MATERIALS AND METHODS: Sida acuta stem segments were placed on PDA medium to isolate endophytic fungi. The fungus was identified by genomic DNA analysis and phylogenetic tree was constructed using ITS sequences (GenBank) to confirm species. The antibacterial efficacy of Aspergillus sulphureus MME12 ethyl acetate extract was tested against Gram-positive and Gram-negative pathogenic bacteria. DPPH free radical scavenging activity, anticancer and DNA fragmentation against EAC cells, and direct haemolytic activity (100-500 µg/mL) using human erythrocytes were determined. RESULTS AND DISCUSSION: The ethyl acetate extract of A. sulphureus (Fresen.) Wehmer (Trichocomaceae) demonstrated significant antibacterial potential against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Salmonella typhi compared to streptomycin. MIC against test pathogens was in the range of 15.6-62.5 µg/mL. The antioxidant results revealed significant RSA from 12.43% to 62.02% (IC50 = 350.4 µg/mL, p ≤ 0.05). MME12 offered considerable inhibition of EAC proliferation (23% to 84%, IC50 = 216.7 µg/mL, p ≤ 0.05) supported by DNA fragmentation studies. The extract also offered insignificant haemolysis (5.6%) compared to Triton X-100. CONCLUSIONS: A single endophytic fungus, A. sulphureus MME12 was isolated and identified using molecular profiling. The above-mentioned findings support the pharmacological application of A. sulphureus MME12 extract and demand for purification of the active principle(s).

Antibacterial and antioxidant properties of biosynthesized zinc oxide nanoparticles from Ceropegia candelabrum L. - An endemic species.

Zinc oxide nanoparticles (ZnO-NPs) were synthesized for the first time from any of the species of Ceropegia. Presently, ZnO-NPs were synthesized from the leaf extract of Ceropegia candelabrum with zinc nitrate using a simple hydrothermal process. The synthesized ZnO-NPs showed an absorption peak at 320nm which is one of the characteristic features of ZnO-NPs. The FT-IR characterization revealed a spectrum band at 551.93cm-1 corresponding to the functional group metal oxide. SEM images showed agglomeration of nanoparticles with a hexagonal shape. XRD results are in corroboration with SEM images as the synthesized particles were of hexagonal wurtzite shape and the size of the particles was in the range of 12-35nm calculated using Scherrer's formula. The elemental analysis using EDS confirmed high zinc content of 70.48% stating that the process of biosynthesis of nanoparticles was carried out in accordance. The biosynthesized ZnO-NPs offered significant antibacterial potential against S. aureus, B. subtilis, E. coli and S. typhi. The antioxidant results revealed significant (p≤0.05) RSA from 0% to 55.43% (IC50=95.09µgmL-1). The results affirm that biosynthesized ZnO-NPs can be used as an alternative to present-day chemical compounds.

Synthesis, Characterization, and BSA Binding Studies of Some New Benzamides Related to Schiff Base.

Condensation of amine 1 with aldehyde 2 gives Schiff base, N-(4-((benzofuran-2-ylmethylene) amino)phenyl)acetamide 3. Schiff base on N-acylation with different substituted acid chlorides in the presence of triethylamine gives the corresponding benzamides, N-acetyl-N-(4-((benzofuran-2-ylmethylene)amino)phenyl)substitutedbenzamide (NABP) 5a-j. The structures of newly synthesized compounds were characterized by elemental analysis, (1)H NMR, (13)C NMR FT-IR, and mass spectral studies. Compounds 3 and 5a-j have been screened for their antimicrobial activity using the disc diffusion and minimum inhibitory concentration (MIC) method against the selected bacterial and fungal strain. Compounds 5a, 5e, 5g, and 5h were found to be more active against all tested strains. The antioxidant properties were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenging methods. Compounds 5i and 5j showed predominant antioxidant activities among the synthesized analogues. The interaction between NABP and bovine serum albumin (BSA) was investigated using fluorescence and ultraviolet spectroscopic techniques at 298 K under imitated physiological conditions. The results revealed that NABP caused the fluorescence quenching of BSA through a static quenching procedure. The binding constants and the number of binding sites were calculated. The binding distance between the donor (BSA) and acceptor (NABP) was determined based on Forster's theory.

Rhizosphere fungus Penicillium chrysogenum promotes growth and induces defence-related genes and downy mildew disease resistance in pearl millet.

Susceptible pearl millet seeds (cv 7042S) were treated with the plant growth promoting fungus Penicillium chrysogenum (PenC-JSB9) at 1 × 10(8) spores·ml(-1) to examine mRNA expression profiles of five defence responsive genes and test its ability to induce resistance to downy mildew caused by Sclerospora graminicola. PenC-JSB9 treatment at 1 × 10(8) CFU·ml(-1) for 6 h significantly enhanced seed germination (9.8- 89%), root length (4.08% to 5.1 cm), shoot length (18.9% to 7.77 cm) and reduced disease incidence (28%) in comparison with untreated controls. In planta colonisation of PenC-JSB9 showed that all three root segments (0-6 cm) and soil dilutions incubated on PDA produced extensive mycelial growth, however colonisation frequency of PenC-JSB9 was significantly higher in soil than in root segments. Spatiotemporal studies revealed that induction of resistance was triggered as early as 24 h and a minimum 2-3 days was optimal for total resistance to build up between inducer treatment and challenge inoculation in both experiments. In Northern blot analysis, transcript accumulation of resistant and PenC-JSB9 induced susceptible cultivars showed higher basal levels of defence gene expression than non-pretreated susceptible controls. Transcript accumulation in resistant seedlings challenge-inoculated with the pathogen showed maximum expression of CHS (3.5-fold increase) and Pr-1a (threefold increase) at 24 and 12 h, respectively. While PenC-JSB9 pretreated susceptible seedlings challenge-inoculated showed rapid and enhanced expression of LOX and POX at 48 h and for CHT at 24 h, whereas non-pretreated susceptible seedlings after pathogen inoculation showed weak expression of hybridised defence genes. Enhanced activation of defence genes by PenC-JSB9 suggests its role in elevated resistance against S. graminicola.

Isolation and characterisation of a protein elicitor from Sclerospora graminicola and elicitor-mediated induction of defence responses in cultured cells of Pennisetum glaucum.

Sclerospora graminicola (Sacc.) Schroet., an oomycete pathogen of Pennisetum glaucum (L.) R.Br. infects the meristematic tissues of young seedlings. The motile zoospores from the sporangia encyst, germinate and penetrate the plant tissue. Resistance to the invading pathogen is governed by the specific recognition of conserved pathogen-associated proteins or elicitors. In the present study, a zoospore protein was isolated and purified to homogeneity by a combination of size exclusion and high-performance liquid chromatography (HPLC). The crude fractionated protein was able to elicit an array of defence responses in resistant and susceptible cells of pearl millet. Treatment of cultured cells of pearl millet with partially purified elicitor protein resulted in a rapid loss of cell viability in the resistant cells and the percentage of cell death was higher in the resistant than in the susceptible cells. Cultures of resistant cells showed a sharp increase in the extra cellular pH compared with susceptible cells when treated with the crude elicitor. Increased oxidative burst was also recorded in the cells treated with the crude elicitor. The purified elicitor showed unique properties. The purified protein was acidic with a pI of 5.6 as revealed by isoelectric focusing (IEF) and matrix-assisted laser desorption ionisation (MALDI) analysis showed that the elicitor had a molecular mass of 7040 daltons. The primary structure determined by N-terminal Edman degradation and searches with BLAST did not reveal similarities to any known plant pathogenic or oomycete elicitor. Higher activities of the important defence-related enzymes phenylalanine ammonia lyase (PAL) and peroxidase in the resistant cell cultures than in the susceptible cell cultures treated with the purified elicitor were clearly evident. Studies of gene expression by northern blotting with heterologus peroxidase, PAL and oxalate oxidase probes showed that the mRNA transcripts were strongly up-regulated in resistant cell cultures within 30 min of elicitor treatment. The purified elicitor also demonstrated a very strong concentration-dependent sterol binding. The purified elicitor protein belongs to a class of low molecular weight oomycete elicitors with sterol carrier properties. The identified low molecular weight protein elicitor displays unique properties that can be exploited for synthesis of novel molecules for eco-friendly crop protection.

Resistance to downy mildew in pearl millet is associated with increased phenylalanine ammonia lyase activity.

Phenylalanine ammonia lyase (PAL) activity was studied in pearl millet cultivars with different levels of resistance to the downy mildew disease caused by Sclerospora graminicola, an important oomycete pathogen. PAL activity was elevated in resistant host cultivar and decreased in susceptible cultivars following downy mildew pathogen infection. The enzyme activation varied between cultivars and was correlated with the degree of resistance to downy mildew disease. The induction of PAL as a response to pathogen inoculation was further corroborated by a time-course study in seedlings and cultured cells of pearl millet. The level of PAL activity was highest at 1.5 h in cultured cells and 4 h in seedlings of resistant host cultivar after inoculation with Sclerospora graminicola. Further studies on PAL activity in different tissues of seedlings showed highest enzyme activity in the young growing region of the root of the resistant host cultivars. The accumulation of wall-bound phenolics and lignin was higher in the resistant cultivar seedlings as evidenced by phloroglucinol-HCl staining and p-coumaric acid assay. The temporal changes in lignin concentration and the concentration of soluble phenolics were greater in root tissues of resistant cultivars than in those of susceptible cultivars. Treatment of resistant seedlings with a PAL inhibitor, α-aminooxy-ß-phenylpropionic acid, resulted in the enhancement of the enzyme activity, whereas in the presence of 1 mm trans-cinnamic acid the pathogen-induced PAL was completely inhibited. Treatment of pearl millet seedlings with exogenously applied PAL inhibitors induced downy mildew disease susceptibility in the resistant pearl millet cultivar, consistent with direct involvement of PAL in downy mildew resistance. Results are discussed with respect to the presumed importance of host phenolic compounds and lignin accumulation and its relation to PAL activation as a response to the pathogen infection.

Induction of Growth Promotion and Resistance Against Downy Mildew on Pearl Millet (Pennisetum glaucum) by Rhizobacteria.

A series of laboratory, greenhouse, and field experiments were conducted to evaluate seven strains of plant growth-promoting rhizobacteria (PGPR). The PGPR were tested as suspensions of fresh cultures and talc-based powder formulations. Evaluations were conducted on pearl millet (Pennisetum glaucum) for growth promotion and management of downy mildew caused by Sclerospora graminicola. All treatments with fresh suspensions and powdered formulations showed enhancement in germination and vigor index over the respective untreated controls. With fresh suspensions, maximum vigor index resulted from treatments by Bacillus pumilus strain INR7 followed by B. subtilis strain IN937b (64 and 38% higher than the untreated control, respectively). With powdered formulation, treatment with strain INR7 also resulted in the highest germination and vigor indexes, which were 10 and 63%, respectively, over the untreated control. Under experimental plot conditions, prominent enhancement in growth also was observed in the disease tests. Yield was enhanced 40 and 37% over the untreated control by seed treatment with powdered formulations of strains INR7 and SE34, respectively. The same strains also increased yield by 36 and 33%, respectively, when applied as fresh suspensions. Studies on downy mildew management resulted in varied degrees of protection by the PGPR both under greenhouse and field conditions. With fresh suspensions, treatment with INR7 resulted in the highest protection (57%), followed by B. pumilus strain SE34 and B. subtilis strain GBO3, which resulted in 50 and 43% protection, respectively, compared with the untreated control. With powdered formulation, PGPR strain INR7 suppressed downy mildew effectively, resulting in 67% protection, while SE34 resulted in 58% protection, followed by GBO3 with 56% protection. Treatment with Apron (Metalaxyl) resulted in the highest protection against downy mildew under both greenhouse and field conditions. Thus, the present study suggests that the tested PGPR, both as powdered formulations and fresh suspensions, can be used within pearl millet downy mildew management strategies and for plant growth promotion.