Management of Strawberry Grey Mold Disease Using Biocontrol Agents and Plant Extracts

Strawberry (Fragaria × ananassa Duch.) is a significant global soft fruit crop, prized for its nutrient content and pleasant flavor. However, diseases, particularly grey mold caused by Botrytis cinerea Pers. Fr. poses major constraints to strawberry production and productivity. Grey mold severely impacts fruit quality and quantity, diminishing market value. This study evaluated five B. cinerea isolates from various locations in the Ri-Bhoi district of Meghalaya. All isolates were pathogenic, with isolate SGM 2 identified as highly virulent. Host range studies showed the pathogen-producing symptoms in the fava bean pods, marigold, gerbera, and chrysanthemum flowers and in the fava bean, gerbera, and lettuce leaves. In vitro tests revealed that neem extract (15% w/v) achieved the highest mycelial growth inhibition at 76.66%, while black turmeric extract (5% w/v) had the lowest inhibition at 9.62%. Dual culture methods with bio-control agents indicated that Bacillus subtilis recorded the highest mean inhibition at 77.03%, while Pseudomonas fluorescens had the lowest at 20.36% against the two virulent isolates. Pot evaluations demonstrated that B. subtilis resulted in the lowest percent disease index at 20.59%, followed by neem extract at 23.31%, with the highest disease index in the control group at 42.51%. Additionally, B. subtilis significantly improved plant growth, yielding an average of 0.32 kg compared to 0.14 kg in the control. The promising results of B. subtilis and neem leaf extract from this study suggest their potential for eco-friendly managing grey mold in strawberries under field conditions.

A glycine-rich nuclear effector VdCE51 of Verticillium dahliae suppresses plant immune responses by inhibiting the accumulation of GhTRXH2

Verticillium dahliae is an important soil-borne fungal pathogen that causes great yield losses in many cash crops.Effectors of this fungus are known to regulate plant immunity but the mechanism much remains unclear.A glycine-rich nuclear effector,VdCE51,was able to suppress immune responses in tobacco against Botrytis cinerea and Sclerotinia sclerotiorum.This effector was a required factor for full virulence of V.dahliae,and its nuclear localization was a requisite for suppressing plant immunity.The thioredoxin GhTRXH2,identified as a positive regulator of plant immunity,was a host target of VdCE51.Our findings show a virulence regulating mechanism whereby the secreted nuclear effector VdCE51 interferes with the transcription of PR genes,and the SA signaling pathway by inhibiting the accumulation of GhTRXH2,thus suppressing plant immunity.

Development of Machine Learning Methods for Accurate Prediction of Plant Disease Resistance

The traditional method of screening plants for disease resistance phenotype is both time-consuming and costly.Genomic selection offers a potential solution to improve efficiency,but accurately predicting plant disease resistance remains a challenge.In this study,we evaluated eight different machine learning(ML)methods,including random forest classification(RFC),support vector classifier(SVC),light gradient boosting machine(lightGBM),random forest classification plus kinship(RFC_K),support vector classification plus kinship(SVC_K),light gradient boosting machine plus kinship(lightGBM_K),deep neural network genomic prediction(DNNGP),and densely connected convolutional networks(DenseNet),for predicting plant disease resistance.Our results demonstrate that the three plus kinship(K)methods developed in this study achieved high prediction accuracy.Specifically,these methods achieved accuracies of up to 95%for rice blast(RB),85%for rice black-streaked dwarf virus(RBSDV),and 85%for rice sheath blight(RSB)when trained and applied to the rice diversity panel I(RDPI).Furthermore,the plus K models performed well in predicting wheat blast(WB)and wheat stripe rust(WSR)diseases,with mean accuracies of up to 90%and 93%,respectively.To assess the generalizability of our models,we applied the trained plus K methods to predict RB disease resistance in an independent population,rice diversity panel II(RDPII).Concurrently,we evaluated the RB resistance of RDPII cultivars using spray inoculation.Comparing the predictions with the spray inoculation results,we found that the accuracy of the plus K methods reached 91%.These findings highlight the effectiveness of the plus K methods(RFC_K,SVC_K,and lightGBM_K)in accurately predicting plant disease resistance for RB,RBSDV,RSB,WB,and WSR.The methods developed in this study not only provide valuable strategies for predicting disease resistance,but also pave the way for using machine learning to streamline genome-based crop breeding.

Utilizing metabolomic approach to study the mode of action of fungicides and corresponding resistance in plant pathogens

Fungicides are an indispensable tool in plant disease control.Various modes of action(MOAs)have been identified in different fungicides to suppress plant pathogens.The combined use of fungicides with distinct MOAs has been recommended to prevent the development of pathogen resistance.In studying MOAs,metabolomics has been proven to be a robust and high-throughput method.Because metabolites are unique and distinct depending on the biological activities of an organism,MOAs can be identified and classified by establishing metabolic fingerprinting and metabolic profiles.Similarly,if fungicide resistance is developed in a pathogen,the metabolome will change,which can be identified.In this review,we have discussed the principles and advanced applications of metabolomics in the study of MOAs and resistance mechanisms of fungicides,and the potential of metabolic data in understanding the interaction between fungicides and pathogens.Challenges are also discussed in the application of metabolomics,improvement of the study on the mechanism of fungicides in their functions against pathogens and advancing the development of novel fungicides.

A major facilitator superfamily transporter PlMFS1 contributes to growth,oosporogenesis,and pathogenesis of Peronophythora litchii

Major facilitator superfamily(MFS)transporters are secondary active membrane transporters that play an important role in solute interchange and energy metabolism.Peronophythora litchii causes the most destructive disease on lichi,litchi downy blight.PlM90 was reported as a key oosporogenesis regulator.Here,we identified an MFS transporter gene PlMFS1,which is up-regulated during oospore formation at the late infection stage,while down-regulated in the PlM90 mutant.To investigate PlMFS1 function,we generated PlMFS1knockout mutants using CRISPR/Cas9-mediated genome editing technology.Compared with the wild-type strain SHS3,PlMFS1 deletion impaired mycelium growth,zoospore release,oospore production and pathogenicity.Furthermore,PlMFS1 deletion significantly affected P.litchii utilization of fructose,lactose and maltose,and may be the PlMFS1 mechanism involved in mycelial growth.PlMFS1 gene deletion also led to deceased laccase activity,laccase-encoding gene downregulation and impaired P.litchii pathogenicity.To our knowledge,this is the first report of an MFS transporter involved in sugar utilization,sexual reproduction,asexual reproduction and pathogenesis in oomycetes.

Constitutive expression of pathogen-inducible OsWRKY31 enhances disease resistance and affects root growth and auxin response in transgenic rice plants

WRKY transcription factors have many regulatory roles we isolated a rice WRKY gene （OsWRKY31） that is induced in response to biotic and abiotic stresses. In this study, by the rice blast fungus Magnaporthe grisea and auxin. This gene encodes a polypeptide of 211 amino-acid residues and belongs to a subgroup of the rice WRKY gene family that probably originated after the divergence of monocot and dicot plants. OsWRKY31 was found to be localized to the nucleus of onion epidermis cells to transiently express OsWRKY31-eGFP fusion protein. Analysis of OsWRKY31 and its mutants fused with a Gal4 DNA-binding domain indicated that OsWRKY31 has transactivation activity in yeast. Overexpression of the OsWRKY31 gene was found to enhance resistance against infection with M. grisea, and the transgenic lines exhibited reduced lateral root formation and elongation compared with wild-type and RNAi plants. The lines with overexpression showed constitutive expression of many defense-related genes, such as PBZI and OsSci2, as well as early auxin-response genes, such as OslAA4 and OsCrll genes. Furthermore, the plants with overexpression were less sensitive to exogenously supplied IBA, NAA and 2,4-1） at high concentrations, suggesting that overexpression of the OsWRKY31 gene might alter the auxin response or transport. These results also suggest that OsWRKY31 might be a common component in the signal transduction pathways of the auxin response and the defense response in rice.

Review Article: High-Temperature Adult-Plant Resistance, Key for Sustainable Control of Stripe Rust

High-temperature adult-plant (HTAP) resistance expresses when plants grow old and the weather becomes warm. This non-race specific and durable type of resistance has been used successfully in control of wheat stripe rust in the US since early 1960s. This article describes practical procedures for identification and characterization of HTAP resistance and reviews recent studies on discovery of genes conferring HTAP resistance. Recent studies providing insights to the molecular basis for the durability of HTAP resistance will be presented. Strategies for improving levels of HTAP resistance and improving control of stripe rust through combining HTAP resistance with effective all-stage resistance will be discussed.

Mapping of QTL conferring leaf rust resistance in Chinese wheat lines W014204 and Fuyu 3 at adult plant stage

Wheat leaf rust is a destructive foliar disease of common wheat （Triticum aestivum L.） worldwide. The most effective, economical s to control the disease is growing resistant cultivars with adult plant resistance （APR）. The Chinese wheat tines W014204 and Fuyu 3 showed high leaf rust resistance in the field. To identify leaf rust APR genes in the two lines, two mapping populations with 215 and 163 F2：3 lines from the crosses W014204/Zhengzhou 5389 and Fuyu 3/Zhengzhou 5389, respectively, were phenotyped for leaf rust severities during the 2010-2011, 2011-2012 and 2012-2013 cropping seasons in the field at Baoding, Hebei Province, China. A total of 1 215 SSR markers were used to identify the quantitative trait loci （QTLs） for leaf rust APR in the two populations. In the W014204/Zhengzhou 5389 population, three QTLs were detected and designated as QLr.hbu-lBL.1, QLr.hbu-2BS.1 and QLr.hbu-7DS, and explained 2.9-8.4, 11.5-38.3 and 8.5-44.5% of the phenotypic variance, respectively; all the resistance alleles at these loci were derived from W014204. In the Fuyu 3/Zhengzhou 5389 population, three QTLs, QLr.hbu-lBL.2, QLr.hbu-2BS.2 and QLr.hbu-7BL, explained 12.0-19.2, 22.3-38.9 and 4.1-4.3% of the phenotypic variance, respectively, and all resistance alleles were contributed by Fuyu 3. Based on chromosome positions of closely linked markers, both QLr.hbu-lBL. 1 and QLr.hbu-lBL.2 are Lr46, and QLr.hbu-7DS is Lr34. QLr.hbu-7BL was mapped on chromosome 7BL near to Lr68 and they are likely the same gene. Based on chromosome positions, pedigree and field reactions, the two 2BS QTLs are different from all the known APR genes and are likely to be newAPR QTL for leaf rust. These QTLs and their closely linked markers are potentially useful for improving leaf rust resistance in wheat breeding.

Characterization of Egyptian <i>Botrytis cinerea</i>Isolates from Different Host Plants

Gray mold causes considerable economic losses of fruit and vegetable production. The current study on Egyptian population structure of Botrytis cinerea demonstrates that this species is composed of four TE genotypes: transposa, vacuma, boty and flipper types using transposable elements and sensitivity to the hydroxyanilide fungicide, fenhexamid. The results show that transposa is the predominant isolate type (63.6%) in the sampled populations of B. cinerea. However, the four isolate types are fenhexamid-sensitive regardless of location, host plant and plant organ. Additionally, B. cinerea isolates collected from different host plants do not exhibit any host preference using artificial infection test on lettuce. Furthermore, no relation is found between isolate type and aggressiveness and no divergence event has occurred among the isolates collected from different locations and host plants. The results suggest that host specialization of B. cinerea has not been occurred in the current sampled crops.

Plant Pathogens Utilize Effectors to Hijack the Host Endoplasmic Reticulum as Part of Their Infection Strategy

1.Introduction As the central organelle in the eukaryotic secretory pathway,the endoplasmic reticulum(ER)mediates cellular processes that include calcium homeostasis and protein processing[1,2].The infection of plants by pathogens can induce ER stress and trigger the unfolded protein response(UPR).The UPR is a conserved protective signaling pathway that leads to programmed cell death(PCD)under extreme conditions[3–5],which can harm or benefit pathogens,depending on the timing and mode of cell death,and on whether the pathogen has physiologically adapted to benefit from the dying tissue[6].The biosynthesis and proper function of plant pattern recognition receptors(PRRs),which perceive pathogen-or microbe-associated molecular patterns(PAMPs or MAMPs)at the cell surface,also rely on N-glycosylation and the ER quality-control(ERQC)system[7–9].However,pathogens have evolved the capacity utilizing effectors to bind to the host ER stress pathway and manipulate it to their advantage during infection.

QTL mapping of adult plant resistance to stripe rust and leaf rust in a Fuyu 3/Zhengzhou 5389 wheat population

Stripe or yellow rust(YR)and leaf rust(LR)cause large losses in wheat production worldwide.Resistant cultivars curtail the levels of losses.The present study aimed to identify quantitative trait loci(QTL)for YR and LR resistance in 147 F2:6 recombinant inbred lines(RIL)derived from the cross Fuyu 3/Zhengzhou 5389.The RIL population and parents were genotyped with the Wheat55 K single nucleotide polymorphism(SNP)array and simple sequence repeat(SSR)markers.All materials were also phenotyped for YR severity at Mianyang in Sichuan province and Baoding in Hebei province in the 2015/2016,2016/2017,and 2017/2018 cropping seasons,and LR severity at Zhoukou in Henan province and at Baoding in 2017/2018.Eleven QTL for YR resistance and five for LR resistance were detected using inclusive composite interval mapping(Ici Mapping).Four of these QTL on chromosomes 1 BL,2 BS,3 AL,and 5 AL conferred resistance to both YR and LR.The QTL on 1 BL was Lr46/Yr29,and that on 7 BL might be Lr68.The QTL on chromosome 2 BS was detected at a similar position to previously detected loci.QYr.hebau-3 AL/QLr.hebau-3 AL,QYr.hebau-5 AL/QLr.hebau-5 AL,QYr.hebau-7 DL,QYr.hebau-4 BS,QYr.hebau-6 DL,and QYr.hebau-2 AS are likely to be new.An SSR marker for QYr.hebau-7 DL was developed and validated in a diverse wheat panel from China,suggesting effectiveness in different genetic backgrounds.These QTL with closely linked SNP and SSR markers could be useful for marker-assisted selection in wheat breeding programs targeting durable resistance to both diseases.

Effect of Plant Extracts on Seed Borne Fungi of Jute

The aim of this study was to determine the prevalence of seed-borne fungal pathogens of jute and their control with plant extract. The experiment was carried out in seed pathology centre, Bangladesh Agricultural University to obtain relevant information about the health of jute seeds and find out the efficacy of extracts as seed-treater. Two varieties viz., Deshipat (Corchorus capsularis) and Tossapat (Corchorus olitorius) were selected for this study and jute seeds were collected from two different locations of Bangladesh. The major predominant identified fungi were Colletotrichum corchori, Macrophomina phaseolina, Fusarium spp., and Botryodiplodia theobromae. Least seed-borne fungi were found in the variety of Tossapat. Low germination of seeds was found related to high prevalence of seed-borne fungal infections. Highest seed-borne infection was obtained in the variety of Deshipat. In this study, we evaluated some major plant extracts such as garlic clove (Allium sativum), neem leaves (Azadirachta indica), allamanda leaves (Allamanda cathartica L.) and marigold leaves (Tagetes spp.) for observing the action of these plant extracts on seed borne fungi of Jute. The study revealed that the extract of Garlic (Allium sativum), and Biskatali (Polygonum hydropiper) increased the germination of seeds and garlic?(Allium sativum) extract was also found to be most effective in controlling the seed-borne infection among all the treatments. Although the inhibiting capacity of the extracts of Allamanda (Allamanda cathartica L.) and Biskatali (Polygonum hydropiper) was found to be similar and Biskatali extract was found to have the second position among all the treated plant extracts in inhibiting the growth of isolated fungal pathogens.

Tetracycline-Based Binary Ti Vectors pLSU with Efficient Cloning by the Gateway Technology for <i>Agrobacterium tumefaciens</i>-Mediated Transformation of Higher Plants

We constructed small high-yielding binary Ti vectors with a bacterial tetracycline resistance gene to facilitate efficient cloning afforded by the Gateway Technology (Invitrogen) for Agrobacterium tumefaciens-mediated transformation of higher plants. The Gateway Technology vectors are kanamycin-based, thus tetracycline-based destination and expression vectors are easily selected for the antibiotic resistance in the Escherichia coli media. We reduced the size of the tetracycline resistance gene TetC from pBR322 to 1468 bp containing 1191 bp of the coding region, 93 bp of 5’-upstream, and 184 bp 3’-downstream region. The final size of binary Ti vector skeleton pLSU11 is 5034 bp. pLSU12 and 13 have the kanamycin resistance NPTII gene as a plant-selectable marker. pLSU13?and 15 contain the hygromycin resistance HPH gene as a selection marker. pLSU13 and 15 also have the β-glucuronidase (GUS) reporter gene in addition to the plant selection marker. We also constructed a mobilizable version of tetracycline-based binary Ti vector pLSU16 in which the mob function of ColE1 replicon was maintained for mobilization of the binary vector from E. coli to A. tumefaciens by tri-parental mating. The final size of binary Ti vector skeleton pLSU16 is 5580 bp. New tetracycline- based binary Ti vectors pLSU12 were found as effective as kanamycin-based vector pLSU2 in promoting a 10-fold increase in fresh weight yield of kanamycin-resistant calli after A. tumefaciens-mediated transformation of tobacco leaf discs. Using the Gateway Technology we introduced the plant-expressible GUSgene to the T-DNA of binary Ti vector pLSU12. Expression of the β-glucuronidase enzyme activity was demonstrated by histochemical staining of the GUS activity in transformed tobacco leaf discs.

Review: Plant Binary Vectors of Ti Plasmid in <i>Agrobacterium tumefaciens</i>with a Broad Host-Range Replicon of pRK2, pRi, pSa or pVS1

This review chronicles the development of the plant binary vectors of Ti plasmid in Agrobacterium tumefaciens during the last 30 years. A binary vector strategy was designed in 1983 to separate the T-DNA region in a small plasmid from the virulence genes in avirulent T-DNA-less Ti plasmid. The small plant vectors with the T-DNA region have been simply now called binary Ti vectors. A binary Ti vector consist of a broad host-range replicon for propagation in A. tumeraciens, an antibiotic resistance gene for bacterial selection and the T-DNA region that would be transferred to the plant genome via the bacterial virulence machinery. The T-DNA region delimited by the right and left border sequences contains an antibiotic resistance gene for plant selection, reporter gene, and/or any genes of interest. The ColEI replicon was also added to the plasmid backbone to enhance the propagation in Escherichia coli. A general trend in the binary vector development has been to increase the plasmid stability during a long co-cultivation period of A. tumefaciens with the target host plant tissues. A second trend is to understand the molecular mechanism of broad host-range replication, and to use it to reduce the size of plasmid for ease in cloning and for higher plasmid yield in E. coli. The broad host-range replicon of VS1 was shown to be a choice of replicon over those of pRK2, pRi and pSA because of the superior stability and of small well-defined replicon. Newly developed plant binary vectors pLSU has the small size of plasmid backbone (4566 bp) consisting of VS1 replicon (2654 bp), ColE1 replicon (715 bp), a bacterial kanamycin (999 bp) or tetracycline resistance gene, and the T-DNA region (152 bp).

In silico Prediction and Analysis of Potential Off-Targets and Off-Target Mutation Detection in StERF3-Gene Edited Potato Plants

The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;however,a few mismatches are tolerated by the Cas9 endonuclease activity.An accurate and careful in silico-based off-target prediction while target selection is preferred to address the issue.These predictions are based on a comprehensive set of selectable parameters.Therefore,we investigated the possible off-target prediction and their screening in StERF3 gene-edited potato plants while developing StERF3-loss-of-function mutants using CRISPR/Cas9 approach.The 201 off-targets for the selected targets of the StERF3 gene were predicted,and 79 werefiltered as potential off-targets.Of these 79,twenty-five off-targets showed scores with defined cut-off values<0.5 and were analyzed in Sterf3-edited potato plants compared to wild-type plants.No off-targeting was found to have occurred in edited plants.

Evaluation of Substrates for Mass Culture of <i>Phaeoacremonium parasiticum</i>, the Fungus Responsible for Artificial Oleoresin Deposition in Agarwood Plant

For establishing the favourable media for growth of Phaeoacremonium parasiticum, the fungus was inoculated in five different media (Potato Dextrose Broth (PDB), Host Extract (50%) + PDB (50%) (HE + PDB), and the solid media viz., Rice Bran (RB), Maize Meal (MM) and Wheat Bran (WB)) at pH of 6.5 and incubated at a temperature of 30&#8451;± 1&#8451;. The solid media were added with water at the rate of 70 ml/100g of solid substrate. After 30 days of inoculation, conidial population of P. parasiticum in liquid media was lower compared to conidial population in solid media. In solid media, MM media showed the highest conidial population (9.56 log&#183;cfu/g) followed by WB (9.50 log&#183;cfu/g) and RB (9.38 log&#183;cfu/g). Hence, Maize Meal media (MM: Water = 100 g:70 ml) at pH of 6.5, incubation temperature: 30&#8451;± 1&#8451;for 30 days would be the standard technique for mass production of P. parasiticum.

Antagonistic Potential of Bacterial Species against Fungal Plant Pathogens(FPP)and Their Role in Plant Growth Promotion(PGP):A Review

Since the 19th century to date,the fungal pathogens have been involved in causing devastating diseases in plants.All types of fungal pathogens have been observed in important agricultural crops that lead to significant pre and postharvest losses.The application of synthetic fungicide against the fungal plant pathogens(FPP)is a traditional management practice but at the same time these fungicides kill other beneficial microbes,insects,animal,and humans and are harmful to environment.The antagonistic microorganism such as bacteria are being used as an alternate strategy to control the FPP.These antagonistic species are cost-effective and eco-friendly in nature.These biocontrol bacteria have a broad mechanism against fungal pathogens present in the phyllosphere and rhizosphere of the plant.The antagonistic bacteria have different strategies against the FPP,by producing siderophore,biofilm,volatile organic compounds(VOCs),through parasitism,antibiosis,competition for limited resources and induce systemic resistance(ISR)in the host plant by activating the immune systems.The commercial bio-products synthesized by the major bacterial species Pseudomonas syringae,Burkholderia cepacia,Streptomyces griseoviridis,Pseudomonas fluorescens and Bacillus subtilis are used to control Fusarium,Pythium,Rhizoctonia,Penicillium,Alternaria,and Geotrichum.The commercial bio-formulations of bacteria act as both antifungal and plant growth regulators.The Plant growth-promoting rhizobacteria(PGPR)played a significant role in improving plant health by nitrogen-fixing,phosphorus solubilization,phytohormones production,minimizing soil metal contamination,and by ACC deaminase antifungal activities.Different articles are available on the specific antifungal activity of bacteria in plant diseases.Therefore,this review article has summarized the information on biocontrol activity of bacteria against the FPP and the role of PGPR in plant growth promotion.This review also provided a complete picture of scattered information regarding antifungal activities of bacteria and the role of PGPR.

Direct Organogenesis from Cotyledonary Node Explants of Cucurbita pepo (L.)—An Important Zucchini Type Vegetable Crop

Cotyledonary node explants from 3 - 5-d-old seedling derived explants of Cucurbita pepo (L.) produced multiple shoots in Murashige and Skoog (MS) salts B5 vitamins containing medium in the presence of N6-benzylamino-purine BAP 1 mg/L + Thidiazuron TDZ 0.05 mg/L. After 4 weeks explants were subcultured to medium containing MS salts B5 Vitamins BAP 0.5 mg/L, Gibberellic acid GA3 1 mg/L and L-Glutamine 15 mg/L. Periodic excision of regenerated shoots from the explants increased shoot efficiency during subculture. The combination of L-Glutamine with BA and GA3 significantly increased the shoot proliferation. Elongated shoots were excised from shoot clumps and transferred to rooting medium containing indole-3-butyric acid (IBA, 1.0 mg/L). The rooted plants were hardened in small pot containing standardized garden soil, well developed plant transferred to greenhouse and survival rate was 85%.

Effect of Planting Density, Irrigation Regimes, and Maize Hybrids with Varying Ear Size on Yield, and Aflatoxin and Fumonisin Contamination Levels

Corn (maize, Zea mays L.) hybrids expressing the flexibility trait in ear size (number of kernels per ear) are marketed for ability to give higher yields under adverse conditions. Altered kernel number is associated with altered number of silk, a major route for infection of kernels by aflatoxin-producing fungi such as Aspergillus flavus. The effect of plant density and irrigation level on yield and accumulation of aflatoxins and fumonisins in harvested grain was compared in a fixed-ear hybrid (Pioneer 33K81), a semi-flexible ear hybrid (Pioneer 3223) and a flexible ear hybrid (Golden Acres 8460) over a range of seeding densities (49,400, 61,750, 74,700, 86,450, and 98,800 seeds·ha–1) in non-irrigated, moderately-irrigated (6.4 cm soil water deficit) and well-irrigated plots (3.8 cm soil water deficit), during three years with variable rainfall. Irrigation increased yields in all hybrids, but in the absence of irrigation, yields were highest with the semi-flexible ear trait hybrid. In general, the hybrid with the flexible ear trait had lower optimal seeding densities than the other hybrids for each soil water regime. In general, kernel number was least affected by seeding density in the hybrid with fixed-ear trait compared to the semi- and flexible ear hybrids. The lowest levels of aflatoxin and of fumonisin contamination in harvested grain were associated with the flexible ear trait at all rainfall and irrigation levels, but there was no evidence that reducing stress by lowering seeding density reduced mycotoxin contamination. Inoculation with A. flavus resulted in much higher levels of aflatoxin and significantly higher levels of fumonisin contamination in grain of all hybrids under most conditions of rainfall and irrigation, suggesting that factors that promote A. flavus infection can affect production of both mycotoxins.

Evaluation of Endophytic Bacteria for Plant Growth Promotion and Pathogen Suppression Traits in Saccharum officinarum

Endophytes are beneficial microbes that are capable of promoting growth, besides protecting colonized plants against plant pathogens. These microbes are of either bacterial, fungal or actinomycetes in plants. In the study, the endophytic bacteria isolated from sugarcane with their characterization related to plant growth promotion and pathogen suppression have been reported. Roots, shoots and leaves of rooted tissue culture plantlets of sugarcane cultivars of 87A298 and 2009A107 were excised aseptically and isolated endophytic bacterial strains. The strains were identified using 16S rRNA gene sequence based homology. Molecular characterization of these strains was carried out for presence of antimicrobial genes. The results showed that the endophytes isolated from sugarcane tissue culture plantlets were of the genera Bacillus （B. amyloliquefaciens, B. subtilis, B. cereus, B. safensis, B. siamensis, B. aryabhattai, B. flexus and B. velezensis） and Paenibacillus pabuli. There were three antimicrobial peptides （AMPs） producing genes of bacilysin, bacillomycin and fengycin in B. amyloliquefaciens （SE1, SE7）, B. siamensis （SE4, SE16）, B. subtilis （SE2, SE3） and B. velezensis （SE15）. The biochemical characterization assays showed that some of these strains could produce hydrogen cyanide （HCN）, protease, cellulase and indole acetic acid （IAA）. Few strains （SE1 and SE4） were phosphate solubilizers, whereas nine isolates were found to be diazotrophs. Most of the bacterial isolates were found antagonistic to Fusarium sacchari, the sugarcane wilt pathogen under in vitro conditions. Overall, the results suggested the scope and potentiality of sugarcane endophytic bacteria, isolated from tissue culture plantlets, in promoting plant growth and suppression of sugarcane pathogen.