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Sugarcane is an important cash crop grown in 137 countries, accounting for 80% of global sugar production. It supports the livelihood of more than 100 million people and up to 25% of the rural population in some countries. Plant-parasitic nematodes are one significant constraint in sugarcane production and can lead to a loss of up to 30% in productivity. More than 300 species of parasitic nematodes have been discovered in sugarcane soil. Owing to limited data, the potential damage to sugarcane crops caused by parasitic nematodes is often underestimated. The main nematodes present in sugarcane fields are root-lesion ( Pratylenchus spp.), spiral ( Helicotylenchus spp.), root-knot ( Meloidogyne spp.), dagger ( Xiphinema spp.), stunt ( Tylenchorhynchus spp.), ring ( Criconemella spp.), and stubby ( Paratrichodoru s spp.). Among these, Meloidogyne javanica and Pratylenchus zeae are the most damaging nematode species. Management of nematodes is a challenging task as there are no clear symptoms of their presence, and they often come in multiple species with varying levels of pathogenicity. Moreover, the management options available are not always effective. Integrated nematode management is a sustainable strategy for controlling nematode infestations. It involves using all possible methods to suppress the parasitic nematode population in a compatible manner and reduce it below economic threshold levels. This article focuses on the challenges of managing nematodes in sugarcane and highlights the opportunity for implementing a sustainable nematode management strategy., Competing Interests: The author(s) declare no conflict of interest.

From the fluff generated during 2005, after the preliminary experiments (2005-2007), a promising clone G2005047 has been identified. It showed moderate resistance to red rot (3.6 on a 9-scale scoring system), less susceptibility to shoot borer (13.25%) and internode borers (25.35%), and resistance to woolly aphid (0%). In the Advanced Yield Trials (2008-2011), it showed advantages over check for cane yield (CY) (11.79%), commercial cane sugar percent (CCSP) (0.35%), and sugar yield (SY) (20.33%). To ascertain its large-scale cultivation suitability, it has experimented under adaptive research trials (2012-2014) at farmers' fields. It exhibited 18.04%, 1.27%, and 19.55% supremacy over the check Co 86032 for CY, CCSP, and SY respectively. The stability of G2005047 under salinity was ascertained through a multi-environment-based experiment (2015-2017). AMMI (Additive Main-effects and Multiplicative Interactions) and GGE (Genotype × Genotype-Environment interaction) biplots were utilized. ANOVA revealed that the genotypic variation exerted the most significant effect followed by genotype × environment interaction and environment. G2005047 had the highest mean values for yield and quality traits with minimal ASV (AMMI stability value) (2.38:CY; 0.57: CCSP; & 0.58:SY) indicating its good-yielding ability and stability. AMMI I, AMMI II, and GGE biplots confirmed the stability of G2005047. In the jaggery quality assessment trials (2018 and 2019), it yielded 37.1% increased jaggery over the check. Also, the clone G2005047, exhibited moderate resistance to red rot disease, less susceptibility to shoot borer (13.25%) and internode borer (25.35%), and resistance against sugarcane woolly aphid (SWA). Due to supremacy for yield, quality, better performance under salinized situations, and tolerance to disease and pests, the clone G2005047 was released as a variety CoG 7 in 2022., (© 2024. The Author(s).)

Background: The Nuclear Factor Y (NF-Y) transcription factor (TF) gene family plays a crucial role in plant development and response to stress. Limited information is available on this gene family in sugarcane., Objectives: To identify sugarcane NF-Y genes through bioinformatic analysis and phylogenetic association and investigate the expression of these genes in response to abiotic and biotic stress., Methods: Sugarcane NF-Y genes were identified using comparative genomics from functionally annotated Poaceae and Arabidopsis species. Quantitative PCR and transcriptome analysis assigned preliminary functional roles to these genes in response to water deficit, cold and African sugarcane borer (Eldana saccharina) infestation., Results: We identify 21 NF-Y genes in sugarcane. Phylogenetic analysis revealed three main branches representing the subunits with potential discrepancies present in the assignment of numerical names of some NF-Y putative orthologs across the different species. Gene expression analysis indicated that three genes, ShNF-YA1, A3 and B3 were upregulated and two genes, NF-YA4 and A7 were downregulated, while three genes were upregulated, ShNF-YB2, B3 and C4, in the plants exposed to water deficit and cold stress, respectively. Functional involvement of NF-Y genes in the biotic stress response were also detected where three genes, ShNF-YA6, A3 and A7 were downregulated in the early resistant (cv. N33) response to Eldana infestation whilst only ShNF-YA6 was downregulated in the susceptible (cv. N11) early response., Conclusions: Our research findings establish a foundation for investigating the function of ShNF-Ys and offer candidate genes for stress-resistant breeding and improvement in sugarcane., (© 2024. The Author(s).)

The continuous utilisation of an alternative host may influence parasitoid performance across successive generations due to conditioning in natal hosts. Tetrastichus howardi (Olliff) has successfully been reared using Tenebrio molitor L. pupae as a feasible alternative host. However, the extended rearing of T. howardi on this alternative host may impact the biological features of the parasitoids. Parasitoids were reared using T. molitor pupae for 30 consecutive generations. Quality criteria were assessed during the generations F5, F15, and F30, offering pupae of the target pest, Diatraea saccharalis (Fabr.), and compared with the F0 generation (parasitoids reared in D. saccharalis pupae). Criteria included assessments of parasitism performance, host selection, and wing form variation in the parasitoid wasps. Additionally, we examined the fecundity of T. howardi females that emerged from both hosts, considering their age, egg loading before and after one oviposition, as well as parasitism of sugarcane stalk borer pupae. Rearing T. howardi using pupae of T. molitor did not affect its biological traits or preference for the target pest for 30 generations. After parasitism, the parasitoid left the host pupa inside the stalk, and one oviposition was enough to kill D. saccharalis pupae and obtain viable parasitoid progeny. Female sexual maturation and egg loading occurred 72 and 96 h after parasitoid emergence. Egg-loading recovery after parasitism did not happen within 24 h. T. howardi can be reared for up to 30 generations using alternative hosts without compromising its parasitism performance or egg loading.

One of the challenges in augmentative biological control programs is the definition of releasing strategy for natural enemies, especially when macro-organisms are involved. Important information about the density of insects to be released and frequency of releases usually requires a great number of experiments, which implies time and space that are not always readily available. In order to provide science-based responses for these questions, computational models offer an in silico option to simulate different biocontrol agent releasing scenarios. This allows decision-makers to focus their efforts to more feasible options. The major insect pest in sugarcane crops is the sugarcane borer Diatraea saccharalis, which can be managed using the egg parasitoid Trichogramma galloi. The current strategy consists in releasing 50,000 insects per hectare for each release, in three weekly releases. Here, we present a simulation model to check whether this releasing strategy is optimal against the sugarcane borer. A sensitive analysis revealed that the population of the pest is more affected by the number of releases rather than by the density of parasitoids released. Only the number of releases demonstrated an ability to drive the population curve of the pest towards a negative growth. For example, releasing a total of 600,000 insects per hectare in three releases led to a lower pest control efficacy that releasing only 250,000 insects per hectare in five releases. A higher number of releases covers a wider range of time, increasing the likelihood of releasing parasitoids at the correct time given that the egg stage is short. Based on these results, it is suggested that, if modifications to the releasing strategy are desired, increasing the number of releases from 3 to 5 at weekly intervals is most likely preferable., (© 2024. The Author(s).)

Purpose: The current most important form of transmission for Trypanosoma cruzi is the oral route, being responsible for high mortality during the acute phase in infected individuals. Thus, this study aimed to investigate the possibility of infection for this parasite using sugarcane juice in different temperatures employing metacyclic trypomastigotes obtained from xenodiagnosis performed in Swiss mice previously infected with T.cruzi Y strain, and then diluted in sugarcane juice., Methods: For stomach histopathological analysis, 20 mice were infected with metacyclic trypomastigotes diluted in sugarcane juice and euthanized after the 2nd, 4th, 10th, and 15th days after infection. Concurrently, six batches of ten mice were fed using 1.5 mL of the mixture and kept for 12 h at the temperatures of - 80 ºC, - 20 ºC, + 2 ºC, + 28 ºC, + 60 ºC, and + 80 ºC, for later infection verification., Results: Inflammatory infiltrate was found after the 2nd day of infection, and amastigotes nests were present after the 4th, 10th, and 15th day in the margo plicatus stomach region. Viable trypomastigotes were observed in the microtubes kept at - 80 ºC, - 20 ºC, and + 2 ºC, but the animal's infection was observed in the - 80 ºC and + 2 ºC groups. In vitro tests demonstrated the decrease of T. cruzi trypomastigote viability, which was negative after 120 h at -20 ºC and 144 h at + 2 ºC, in contrast to the maintenance of survival after 168 h at - 80 ºC., Conclusion: We observed the ability of survival and infection of T. cruzi packaged at - 80 ºC without the use of preservatives and, therefore, less suitable for storing food., (© 2023. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Native Australian soldier flies, Inopus spp. (Diptera: Stratiomyidae), are agricultural pests of economic importance to the sugarcane industry. A screen of the salivary gland transcriptome of Inopus flavus (James) revealed the presence of viral RNA belonging to a potentially novel member of the family Dicistroviridae. The complete genome sequence consists of 9793 nucleotides with two open reading frames. The genome includes two potential internal ribosomal entry sites (IRESs): one within the 5' UTR and the other in the intergenic region (IGR). Virus particles purified from infected larvae and visualised by electron microscopy were found to be icosahedral, non-enveloped, and 30 nm in diameter., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Sugarcane (Saccharum officinarum), a sugar crop commonly grown for sugar production all over the world, is susceptible to several insect pests attack in addition to bacterial, fungal and viral infections leading to substantial reductions in its yield. The complex genetic makeup and lack of resistant genes in genome of sugarcane have made the conventional breeding a difficult and challenging task for breeders. Using pesticides for control of the attacking insects can harm beneficial insects, human and other animals and the environment as well. As alternative and effective strategy for control of insect pests, genetic engineering has been applied for overexpression of cry proteins, vegetative insecticidal proteins (vip), lectins and proteinase inhibitors (PI). In addition, the latest biotechnological tools such as host-induced gene silencing (HIGS) and CRISPR/Cas9 can be employed for sustainable control of insect pests in sugarcane. In this review overexpression of the cry, vip, lectins and PI genes in transgenic sugarcane and their disease resistance potential is described.

Sugarcane (Saccharum officinarum L.) is a cash crop grown commercially for its higher amounts of sucrose, stored within the mature internodes of the stem. Numerous studies have been done for the resistance development against biotic and abiotic stresses to save the sucrose yields. Quality and yield of sugarcane production is always threatened by the damages of cane borers and weeds. In current study two problems were better addressed through the genetic modification of sugarcane for provision of resistance against insects and weedicide via the expression of two modified cane borer resistant CEMB-Cry1Ac (1.8 kb), CEMB-Cry2A (1.9 kb) and one glyphosate tolerant CEMB-GTGene (1.4 kb) genes, driven by maize Ubiquitin Promoter and nos terminator. Insect Bio-toxicity assays were carried out for the assessment of Cry proteins through mortality percent of shoot borer Chilo infuscatellus at 2nd instar larvae stage. During V 0 , V 1 and V 2 generations young leaves from the transgenic sugarcane plants were collected at plant age of 20, 40, 60, 80 days and fed to the Chilo infuscatellus larvae. Up to 100% mortality of Chilo infuscatellus from 80 days old transgenic plants of V 2 generation indicated that these transgenic plants were highly resistant against shoot borer and the gene expression level is sufficient to provide complete resistance against target pests. Glyphosate spray assay was carried out for complete removal of weeds. In V 1 -generation, 70-76% transgenic sugarcane plants were found tolerant against glyphosate spray (3000 mL/ha) under field conditions. While in V 2 -generation, the replicates of five selected lines 4L/2, 5L/5, 6L/5, L8/4, and L9/6 were found 100% tolerant against 3000 mL/ha glyphosate spray. It is evident from current study that CEMB-GTGene, CEMB-Cry1Ac and CEMB-Cry2A genes expression in sugarcane variety CPF-246 showed an efficient resistance against cane borers (Chilo infuscatellus) and was also highly tolerant against glyphosate spray. The selected transgenic sugarcane lines showed sustainable resistance against cane borer and glyphosate spray can be further exploited at farmer's field level after fulfilling the biosafety requirements to boost the sugarcane production in the country.

Seasonal temperature and precipitation patterns on a global scale are main factors to which insects and plants adapt through natural selection, although periodic outbreaks in insect populations may occur in areas where they had not been previously reported, a phenomenon considered as a consequence of global warming. In this study, we estimate the distribution of sugarcane borers, Diatraea spp., under different climate scenarios (rcp26, rcp45, rcp60 and rcp85.) Insects were collected weekly in four sugarcane fields from four different towns in the department of Caldas (Colombia) during 2017, and also in several sugarcane fields in the Cauca River Valley (CRV) between 2010 and 2017. The influence of climatic variables on different agro-ecological zones of the CRV sugarcane fields was defined by climatic data between 2010 and 2017 (maximum and minimum daily temperatures, and accumulated precipitation). The estimate of an optimal niche for Diatraea spp. includes temperatures between 20°C and 23°C, accumulated annual rainfall between 1200 and 1500 mm, dry months with precipitations below 50 mm, slopes of less than 0.05 degrees, crop heterogeneity with an index of 0.2 and primary production values of 1.0. Data suggests Diatraea population is considerably influenced by adverse climate change effects, under the premise of an increase in local and global temperatures, reducing its population niches as well as the number of individuals., Competing Interests: The authors have declared that no competing interests exist.

Diatraea saccharalis constitutes a threat to the sugarcane productivity, and obtaining borer tolerant cultivars is an alternative method of control. Although there are studies about the relationship between the interaction of D. saccharalis with sugarcane, little is known about the molecular and genomic basis of defense mechanisms that confer tolerance to sugarcane cultivars. Here, we analyzed the transcriptional profile of two sugarcane cultivars in response to borer attack, RB867515 and SP80-3280, which are considered tolerant and sensitive to the borer attack, respectively. A sugarcane genome and transcriptome were used for read mapping. Differentially expressed transcripts and genes were identified and termed to as DETs and DEGs, according to the sugarcane database adopted. A total of 745 DETs and 416 DEGs were identified (log 2 |ratio| > 0.81; FDR corrected P value ≤ 0.01) after borer infestation. Following annotation of up- and down-regulated DETs and DEGs by similarity searches, the sugarcane cultivars demonstrated an up-regulation of jasmonic acid (JA), ethylene (ET), and defense protein genes, as well as a down-regulation of pathways involved in photosynthesis and energy metabolism. The expression analysis also highlighted that RB867515 cultivar is possibly more transcriptionally activated after 12 h from infestation than SP80-3280, which could imply in quicker responses by probably triggering more defense-related genes and mediating metabolic pathways to cope with borer attack.

The giant sugarcane borer Telchin licus (Drury, 1773) (Lepidoptera: Castniidae) is a day-flying moth pest of sugarcane, pineapples and bananas. To better understand the chemical communication in this species, we examined the morphology of its olfactory system and the chemical composition of its body parts. The ventral surface of the clubbed antennae of T. licus has six morphological types of sensilla: sensilla trichodea, basiconica, chaetica, squamiforma, coeloconica, and auricillica. The telescopic ovipositor shows no evidence of a sexual gland, or female-specific compounds. On the other hand, the midleg basitarsus of males releases (E,Z)-2,13-octadecadienol and (Z,E)-2,13-octadecadienol, which are electroantennographically active in both sexes. These compounds are known female sex pheromones in the Sesiidae family and are male-specific compounds in another castniid moth, although further investigations are necessary to elucidate their ecological role in the Castniidae family., Competing Interests: The authors have declared that no competing interests exist.

The recognition of chemical signal including volatile odorants and pheromones is very important in the olfactory physiological behaviors of insects, such as avoiding predators, seeking food and mating partners. The sugarcane borer, Tryporyza intacta is the most harmful insect in sugarcane region in Southeast Asia and Southern China, however, the study of their molecular biology and physiology was limited. Here we demonstrated that the sex pheromone (E11-16:Ald: Z11-16:Ald = 7:3) were most effective to T. intacta. In addition, compared the traditional rubber lure, a new microsphere formulation lure can optimize the trapping effect and might be widely used in the sugarcane growing area. To obtain a better understanding of the olfactory molecular mechanism of pheromone-based mate recognition system, we have cloned the full-length gene of the TintPBP4 and expressed in Escherichia coli. Our phylogenetic analysis highlighted that the TintPBP4 was highly conserved among diverse species of Lepidoptera. Furthermore, the results of QRT-PCR demonstrated that TintPBP4 transcripts were abundantly expressed in the antennae of T. intacta, especially in the male adults. The fluorescence binding experiments showed the TintPBP4 exhibited strong binding capacities to the sex pheromone components. These results will not only provide more understanding for the functional analysis of olfactory proteins from T. intacta, but also assist in the exploitation and development of sex pheromones in the integrated biological control of this pest.

Introduction: Oral transmission of acute Chagas disease is an emerging public health concern. This study aimed to detect insect fragments in experimentally contaminated food, by comparing triatomines with other insects., Methods: Food samples were experimentally contaminated with insects, processed to recover their fragments by light filth, and analyzed by microscopy and Polymerase Chain Reaction (PCR)., Results: Morphological differences between coleopteran and triatomine insects were observed in microscopic images. PCR was efficient in amplifying Triatominae DNA in the experimentally contaminated food., Conclusions: This methodology could be utilized by food analysts to identify possible insect contamination in food samples.

Root-lesion nematode ( Pratylenchus zeae ) and root-knot nematode ( Meloidogyne javanica ) are two important pathogens of sugarcane ( Saccharum hybrid). No commercial cultivars are resistant to these nematodes in Australia. Twenty accession lines of S. spontaneum , a wild relative of sugarcane, were tested against these two nematode species. S. spontaneum lines were tested twice for resistance to root-lesion nematode and three times for root-knot nematode. Reproduction (final population/starting population) of root-lesion nematodes was significantly lower in 17 of the 20 S. spontaneum accession lines tested in two experiments compared with two commercial cultivars. Four S. spontaneum lines supported a significantly lower number of root-lesion nematodes per gram of root than that of two commercial sugarcane cultivars. Reproduction of root-knot nematodes was significantly lower in 16 S. spontaneum lines compared with two commercial cultivars. Fourteen of the S. spontaneum lines tested supported significantly fewer eggs per gram of root compared with two commercial cultivars. This study showed that S. spontaneum lines possessed resistance for root-lesion and root-knot nematodes. Targeted crossing with commercial hybrid parental lines should be conducted to introduce nematode resistance into sugarcane cultivars for the Australian sugar industry.

The aim of this work was to use spectroscopic methods and partial least squares discriminant analysis (PLS-DA) for the early prediction of genotype resistance or susceptibility to sugarcane borer. The sugarcane leaf +1 was directly analyzed with no sample preparation by ultraviolet-visible-near-infrared (UV-VIS-NIR), middle-infrared (MID), and near-infrared (NIR) spectroscopies. Also, laser-induced breakdown spectroscopy (LIBS) was used to analyze pellets of dried and ground leaves and stalks of sugarcane. Classification models were built using PLS-DA. The models built using UV-VIS-NIR, MID or NIR spectra exhibited ideal sensitivity, specificity, and classification errors, i.e., 1 for both sensitivity and specificity and 0 for classification errors. Regarding the models built using LIBS spectra, those using spectra of pellets made from dried and ground leaves also presented ideal sensitivity, specificity, and classification errors; on the other hand, models built using the spectra of pellets made of dried and ground stalks did not present ideal values for these parameters. Thus, the models built, except for the one using LIBS of pellets made of stalks, showed excellent predictive capacity, making them suitable for predicting the resistance or susceptibility of sugarcane genotypes in the early stages of a plant's life., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Sugarcane (Saccharum spp. hybrids) is considered the most globally important sugar-producing crop and raw material for biofuel. Insect attack is a major issue in sugarcane cultivation, resulting in yield losses and sucrose content reductions. Stem borer (Diatraea saccharalis F.) causes serious yield losses in sugarcane worldwide. However, insect-resistant germplasms for sugarcane are not available in any collections all over the world, and the molecular mechanism of insect resistance has not been elucidated. In this study, cry1Ac transgenic sugarcane lines were obtained and the biological characteristics and transgene dosage effect were investigated and a global exploration of gene expression by transcriptome analysis was performed., Results: The transgene copies of foreign cry1Ac were variable and random. The correlation between the cry1Ac protein and cry1Ac gene copies differed between the transgenic lines from FN15 and ROC22. The medium copy lines from FN15 showed a significant linear relationship, while ROC22 showed no definite dosage effect. The transgenic lines with medium copies of cry1Ac showed an elite phenotype. Transcriptome analysis by RNA sequencing indicated that up/down regulated differentially expressed genes were abundant among the cry1Ac sugarcane lines and the receptor variety. Foreign cry1Ac gene and endogenous borer stress-related genes may have a synergistic effect. Three lines, namely, A1, A5, and A6, were selected for their excellent stem borer resistance and phenotypic traits and are expected to be used directly as cultivars or crossing parents for sugarcane borer resistance breeding., Conclusions: Cry1Ac gene integration dramatically improved sugarcane insect resistance. The elite transgenic offspring contained medium transgene copies. Foreign cry1Ac gene integration and endogenous borer stress-related genes may have a synergistic effect on sugarcane insect resistance improvement.

The purpose of this study was to investigate and compare root morpho-anatomical traits and physiological responses of susceptible (SP81-3250) and resistant (H. Kawandang) sugarcane genotypes exposed to the attack by nymphs of spittlebug Mahanarva fimbriolata (Stål) (Hemiptera: Cercopidae). Two experiments were conducted to compare the damage caused by spittlebug nymphs on fresh and dry biomass weight; lignin content in stalks; root anatomy; chlorophyll content; photosynthetic rate (A); carboxylation efficiency (A/Ci); stomatal conductance (gS) and transpiration rate (E) of these genotypes. SP81-3250 consistently obtained significantly higher damage scores than H. Kawandang in both experiments, confirming the previously observed level of resistance in each genotype. Attack by spittlebug nymphs had a much higher effect on both fresh and dry biomass weight, chlorophyll content, A, A/Ci, gs and E of SP81-3250, than that on H. Kawandang. Anatomical studies indicated the presence of aerenchyma tissue in the root cortex of SP81-3250, a feature which may facilitate penetration of the nymph's stylet into the vascular cylinder. In contrast, roots of H. Kawandang are characterized by having more dense and compact parenchyma cells. In addition, infested plants of this genotype contained an unidentified mucilaginous compound in the vascular cylinder of the roots. We conclude that resistance of H. Kawandang to spittlebug is related to the ability of this genotype to maintain normal chlorophyll content, as well as stomatal conductance and photosynthesis, thus, allowing for biomass accumulation under spittlebug attack, in contrast to SP81-3250. In addition, the presence of more compact and denser parenchymal cells, as well as that of an induced mucilaginous compound in the root's vascular cylinder, are likely to hinder host-feeding activity in nymphs, causing higher nymph mortality and therefore, reduced damage in plants of this genotype.

In this study, we investigated resistance traits to the sugarcane borer Diatraea saccharalis Fab. (Lepidoptera: Crambidae) in the leaves and stalks of six sugarcane cultivars in a series of greenhouse and laboratory assays. Investigation of plant factors and infestation rates to better discriminate stalk damage by the sugarcane borer indicated that infestation of 7-month-old, single plants with 20 larvae at the third or fourth instar per plant was suitable to assess tunneling length. Three cultivars (i.e. SP803280, RB928064, and RB835486) had lower stalk damage (i.e. tunnel length) than cultivar SP891115, which exhibited relatively greater susceptibility to tunneling by the borer. The time required for the larvae to enter the sugarcane stalk was longer for cultivar SP803280, indicating resistance traits on the stalk surface, which correlated with lower stalk damage. Larvae feeding on SP813250 stalks had the lowest weight gain, indicating that this cultivar has resistance traits to larval development within its stalks. Cultivars RB867515 and SP891115 resulted in the highest mortality of early-stage larvae feeding on leaves, indicating the presence of resistance factors in their leaves. Multi-trait cluster and principal component analyses placed the cultivars into three and four clusters, respectively. The cultivars placed in different groups that exhibited resistance to leaf feeding, stalk entrance, and tunneling by the sugarcane borer could be used for crossings in sugarcane breeding programs with the goal of obtaining higher levels of resistance to D. saccharalis.

Sugarcane borer is the most common and harmful pest in Chinese sugarcane fields, and can cause damage to the whole plant during the entire growing season. To improve borer resistance in sugarcane, we constructed a plant expression vector pGcry2A0229 with the bar gene as the marker and the cry2A gene as the target, and introduced it into embryogenic calli of most widely cultivated sugarcane cultivar ROC22 by particle bombardment. After screening with phosphinothricin in vitro and Basta spray, 21 resistance-regenerated plants were obtained, and 10 positive transgenic lines harboring the cry2A gene were further confirmed by conventional PCR detection. Real-time quantitative PCR (RT-qPCR) analysis showed that the copy number of the cry2A gene varied among different transgenic lines but did not exceed four copies. Quantitative ELISA analysis showed that there was no linear relationship with copy number but negatively correlated with the percentage of borer-infested plants. The analysis of industrial and agronomic traits showed that the theoretical sugar yields of transgenic lines TR-4 and TR-10 were slightly lower than that of the control in both plant cane and ratoon cane; nevertheless, TR-4 and TR-10 lines exhibited markedly lower in frequency of borer-infested plants in plant cane and in the ratoon cane compared to the control. Our results indicate that the introduction of the cry2A gene via bombardment produces transgenic lines with obviously increased stem borer resistance and comparable sugar yield, providing a practical value in direct commercial cultivation and crossbreeding for ROC22 has been used as the most popular elite genitor in various breeding programs in China.

A cercomonad, named Phytocercomonas venanatans, is confirmed as the cause of the sugarcane disease chlorotic streak. This was achieved by establishing aseptic liquid cultures of the pathogen isolated from internal pieces of sugarcane stalk tissue. Actively motile cultures of the pathogen were inoculated into sugarcane roots, stalks, and leaf whorls. Infected plants subsequently developed the characteristic symptoms of chlorotic streak. Infection was confirmed by PCR screening of plant tissues and by reisolation of the pathogen into aseptic culture followed by PCR and microscopic confirmation. P. venanatans is the first reported pathogenic cercomonad able to systemically infect higher plants and the first plant pathogenic cercozoan able to be successfully grown in axenic culture on common microbiological media.

Chlorotic streak is a global disease of commercial sugarcane (Saccharum spp. hybrids). The disease is transmitted by wet soil, water, as well as in diseased planting material. Although first recognized almost 90 years ago and despite significant research effort, the identity of the causal agent has been elusive. Metagenomic high throughput sequencing (HTS) facilitated the discovery of novel protistan ribosomal and nuclear genes in chlorotic streak-infected sugarcane. These sequences suggest a possible causal agent belonging to the order Cercomonadida (Rhizaria, phylum Cercozoa). An organism with morphological features similar to cercomonads (=Cercomonadida) was isolated into pure axenic culture from internal stalk tissues of infected sugarcane. The isolated organism contained DNA sequences identical to those identified in infected plants by HTS. The DNA sequences and the morphology of the organism did not match any known species. Here we present a new genus and species, Phytocercomonas venanatans, which is associated with chlorotic streak of sugarcane. Amplicon sequencing also supports that P. venanatans is associated with this disease. This is the first reported member from Cercomonadida showing a probable pathogenic association with higher plants.

The aim of this article was to confirm and describe an outbreak of acute Chagas disease involving oral transmission in the western region of Rio Grande do Norte State, Brazil. This was a descriptive case series study in which the data sources were medical records and interviews with suspected cases from September 16 to November 19, 2015. An entomological investigation was conducted in the probable sites of infection for acute Chagas disease cases. Eighteen cases of acute Chagas disease were confirmed in residents of four municipalities (counties) in Rio Grande do Norte State. The most frequently reported signs and symptoms were fever and weakness (n = 18), followed by myalgia (n = 17), prostration, loss of appetite, and edema of the lower limbs (n = 15). Median duration of fever was 20 days (range: 6 to 45 days). Fifteen cases were confirmed by the laboratory criterion and three by epidemiological link with consistent clinical characteristics. All confirmed cases reported having consumed sugar cane juice from the same mill. A total of 110 triatomines were captured on the plantation where the sugar cane had been crushed for juice. The insects were found in the peridomicile, in stacks of firewood and close to the sugar cane mill. The majority of the captured specimens were Triatoma brasiliensis and showed a natural infection rate of 63%. The Chagas disease outbreak was confirmed with oral transmission via ingestion of sugar cane juice contaminated with infected triatomines, as evidenced by the epidemiological link between the investigated cases and the entomological survey in the probable site where the infection occurred.

Human-mediated changes in landscapes can facilitate niche expansion and accelerate the adaptation of insect species. The interaction between the evolutionary history of the sugarcane borer, Diatraea saccharalis Fabricius, and historical and modern agricultural activity in Brazil shaped its spatial genetic structure, facilitating ecological divergence and incipient host shifting. Based on microsatellite data, STRUCTURE analyses identified two (K = 2) and three (K = 3) significant genetic clusters that corresponded to: (a) a strong signal of spatial genetic structure and, (b) a cryptic signal of host differentiation. We inferred that K = 2 reflects the footprint of agricultural activity, such as expansion of crop production (sugarcane and maize), unintentional dispersion of pests, and management practices. In contrast, K = 3 indicated incipient host differentiation between larvae collected from sugarcane or maize. Our estimates of population size changes indicated that a historical bottleneck was associated with a reduction of sugarcane production ≈200 years ago. However, a more recent population expansion was detected (>1950s), associated with agricultural expansion of large crop production into previously unfarmed land. Partial Mantel tests supported our hypothesis of incipient host adaptation, and identified isolation-by-environment (e.g., host plant) in São Paulo and Minas Gerais states, where sugarcane has been traditionally produced in Brazil. The impact of agricultural production on D. saccharalis may continue, as the current population structure may hinder the efficacy of refuge plants in delaying insect resistance evolution to Bt toxin.

Chagas disease, caused by Trypanosoma cruzi affects about 6-8 million people worldwide. Although transmission by triatomine insects has been controlled, other means of transmission maintain the infection. These forms of transmission are responsible for introducing Chagas disease in other non-endemic countries of the world. Thus, Chagas disease, nowadays is a worldwide health problem. In Brazil, acai pulp and sugarcane juice have been associated with Chagas disease outbreaks. The difficulties in isolation of the parasite from foods are hampering source tracking which could allow the confirmation of an implicated food commodity in these outbreak investigations. To address this scientific gap, we evaluated the performance of real-time PCR (qPCR) for detecting T. cruzi in acai pulp and sugarcane juice. All experiments were performed with acai pulp and sugarcane juice samples contaminated with different concentrations of T. cruzi. In assays with qPCR, the results showed that the ideal procedure for T. cruzi identification in acai pulp and sugarcane juice consisted of: i. centrifugation; ii. DNA extraction with a commercial kit for stool matrix; and iii. qPCR using a specific molecular marker for T. cruzi. The seeding in LIT medium of experimentally contaminated foods was effective in detecting the parasitic load by qPCR. The efficacy of qPCR was also verified testing food samples crushed with infected Triatomines. In conclusion, this methodology can be used to perform rapid diagnosis in outbreaks, facilitating measures in disease control., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: The red palm weevil (RPW), Rhynchophorus ferrugineus, is an invasive wood-boring insect that damages palms and sugarcane. Bacillus thuringiensis (Bt) is an entomopathogenic bacterium which has been modified into various strains and widely used in pest management. The aim of this study was to evaluate the susceptibility of RPW to the HA strain of Bt., Results: Five concentrations of Bt bioassays were used on RPW eggs, second instars and fourth instars. Average egg hatching rates exceeded 85% using Bt suspensions or distilled water. Hatch times were extended significantly using higher Bt concentrations. For second instar larvae, the LC 50 was 4.92 × 10 9 CFU mL -1 15 d after feeding; the LT 50 values decreased with each higher concentration. The corrected mortality of second instars increased significantly with increased concentrations after 15 d, ranging from 16.97% to 94.32%. Significant differences occurred in the boring activity of fourth instars when dipped in Bt suspensions or crawling on treated sugarcane. Bacterial infection in dead larvae was confirmed using molecular techniques., Conclusion: Our results indicated that Bt can be used in RPW control as a potential biological control agent and can effectively reduce palm trees damage. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Sugarcane, a major crop grown in the tropical and subtropical areas of the world, is produced mainly for sucrose, which is used as a sweetener or for the production of bioethanol. Among the numerous pests that significantly affect the yield of sugarcane, the sugarcane rhizome borer (Migdolus fryanus, a cerambycidae beetle) is known to cause severe damage to the crops in Brazil. The absence of molecular information about this insect reinforces the need for studies and an effective method to control this pest. In this study, RNA-Seq technology was employed to study different parts of M. fryanus larvae. The generated data will help in further investigations about the taxonomy, development, and adaptation of this insect. RNA was extracted from six different parts (head, fat body, integument, hindgut, midgut, and foregut) using Trizol methodology. Using Illumina paired-end sequencing technology and the Trinity platform, trimming and de novo assembly was performed, resulting in 44,567 contigs longer than 200 nt for a reunion of data from all transcriptomes, with a mean length of 1,095.27 nt. Transcripts were annotated using BLAST against different protein databanks (Uniprot/Swissprot, PFAM, KEEG, SignalP 4.1, Gene Ontology, and CAZY) and were compared for similarity using a Venn diagram. Differential expression patterns were studied for select genes through qPCR and FPKM comprising important protein families (digestive peptidases, glucosyl hydrolases, serine protease inhibitors and otopetrin), which allowed a better understanding of the insect's digestion, immunity and gravity sensorial mechanisms.

Schizaphis graminum (green bug; GB) and Sipha flava (yellow sugarcane aphid; YSA) are two cereal aphid species with broad host ranges capable of establishing on sorghum (Sorghum bicolor) and several switchgrass (Panicum virgatum) cultivars. Switchgrass and sorghum are staple renewable bioenergy crops that are vulnerable to damage by aphids, therefore, identifying novel targets to control aphids has the potential to drastically improve yields and reduce losses in these bioenergy crops. Despite the wealth of genomic and transcriptomic information available from a closely related model aphid species, the pea aphid (Acyrthosiphon pisum), similar genomic information, including the identification of small RNAs, is still limited for GB and YSA. Deep sequencing of miRNAs expressed in GB and YSA was conducted and 72 and 56 miRNA candidates (including 14 and eight novel) were identified, respectively. Of the identified miRNAs, 45 were commonly expressed in both aphid species. Further, plant derived miRNAs were also detected in both aphid samples, including 13 (eight known and five novel) sorghum miRNAs and three (novel) barley miRNAs. In addition, potential aphid gene targets for the host plant-derived miRNAs were predicted. The establishment of miRNA repertoires in these two aphid species and the detection of plant-derived miRNA in aphids will ultimately lead to a better understanding of the role of miRNAs in regulating gene expression networks in these two aphids and the potential roles of plant miRNAs in mediating plant-insect interactions., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Sugarcane's (Saccharum spp.) response to Diatraea saccharalis (F.) (Lepidoptera: (Crambidae) herbivory was investigated using a macroarray spotted with 248 sugarcane Expressed Sequence Tags (ESTs) encoding serine peptidase inhibitors, serine peptidases. and Clp protease system subunits. Our results showed that after nine hours of herbivory, 13 sugarcane genes were upregulated and nine were downregulated. Among the upregulated genes, nine were similar to serine peptidase inhibitors and four were similar to Bowman-Birk Inhibitors (BBIs). Phylogenetic analysis revealed that these sequences belong to a phylogenetic group of sugarcane BBIs that are potentially involved in plant defense against insect predation. The remaining four upregulated genes included serine peptidases and one homolog to the Arabidopsis AAA+ chaperone subunit ClpD, which is a member of the Clp protease system. Among the downregulated genes, five were homologous to serine peptidases and four were homologous to Arabidopsis Clp subunits (three homologous to Clp AAA+ chaperones and one to a ClpP-related ClpR subunit). Although the roles of serine peptidase inhibitors in plant defenses against herbivory have been extensively investigated, the roles of plant serine peptidases and the Clp protease system represent a new and underexplored field of study. The up- and downregulated D. saccharalis genes presented in this study may be candidate genes for the further investigation of the sugarcane response to herbivory., Competing Interests: The authors declare no conflicts of interest.

The Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae), is an invasive pest of sugarcane, Saccharum spp., rice, Oryza sativa L., and other graminaceous crops in the United States. Traps baited with the synthetic female sex pheromone of E. loftini are used for monitoring and management of this invasive pest. However, the active space, or radius of attraction, of these traps is not known. Two field experiments examined the effect of intertrap distance on trap captures with hexagonal arrays of traps deployed in rice stubble habitat in Texas (2011) and Louisiana (2013). Trap capture increased with increasing intertrap distance. Trap interference occurred at intertrap distances ≤50 m in the 2011 experiment. Results from the experiment conducted in 2013 indicate that trap interference occurs at intertrap distances of 50 m, but not at distances ≥100 m. These results suggest that under field conditions, E. loftini pheromone traps attract males from distances of 50-100 m. The active space of pheromone traps also was examined under controlled wind conditions by direct observation of male response to detection of the female sex pheromone. Eoreuma loftini males responded to the pheromone blend by becoming active, fanning their wings, and rapidly walking in circles. The mean distance from the pheromone source at which males responded was 47.6 m. This work provides the first documentation of active space for traps baited with female sex pheromone for a crambid species, and these data will improve pheromone trap deployment strategies for E. loftini monitoring and management.

We developed sugarcane plants with improved resistance to the sugarcane borer, Diatraea saccharalis (F). An expression vector pGcry1Ac0229, harboring the cry1Ac gene and the selectable marker gene, bar, was constructed. This construct was introduced into the sugarcane cultivar FN15 by particle bombardment. Transformed plantlets were identified after selection with Phosphinothricin (PPT) and Basta. Plantlets were then screened by PCR based on the presence of cry1Ac and 14 cry1Ac positive plantlets were identified. Real-time quantitative PCR (RT-qPCR) revealed that the copy number of cry1Ac gene in the transgenic lines varied from 1 to 148. ELISA analysis showed that Cry1Ac protein levels in 7 transgenic lines ranged from 0.85 μg/FWg to 70.92 μg/FWg in leaves and 0.04 μg/FWg to 7.22 μg/FWg in stems, and negatively correlated to the rate of insect damage that ranged from 36.67% to 13.33%, respectively. Agronomic traits of six transgenic sugarcane lines with medium copy numbers were similar to the non-transgenic parental line. However, phenotype was poor in lines with high or low copy numbers. Compared to the non-transgenic control plants, all transgenic lines with medium copy numbers had relatively equal or lower sucrose yield and significantly improved sugarcane borer resistance, which lowered susceptibility to damage by insects. This suggests that the transgenic sugarcane lines harboring medium copy numbers of the cry1Ac gene may have significantly higher resistance to sugarcane borer but the sugarcane yield in these lines is similar to the non-transgenic control thus making them superior to the control lines.

Metridius nov. gen. and types species M. robertsoni nov. sp. with winged males and apterous females is described from adults and larvae found infesting sugarcane stools from near Mackay, central Queensland. A second new species, M. mcalpinei nov. sp., based only on males from near Sydney, New South Wales is also described. Notes on the biology of both species and an identification key to the genera of the subfamily Chiromyzinae and to the species are also given.

Most aphids are highly specialized on one or two related plant species and generalist species often include sympatric populations adapted to different host plants. Our aim was to test the hypothesis of the existence of host specialized lineages of the aphid Melanaphis sacchari in Reunion Island. To this end, we investigated the genetic diversity of the aphid and its association with host plants by analyzing the effect of wild sorghum Sorghum bicolor subsp. verticilliflorum or sugarcane as host plants on the genetic structuring of populations and by performing laboratory host transfer experiments to detect trade-offs in host use. Genotyping of 31 samples with 10 microsatellite loci enabled identification of 13 multilocus genotypes (MLG). Three of these, Ms11, Ms16 and Ms15, were the most frequent ones. The genetic structure of the populations was linked to the host plants. Ms11 and Ms16 were significantly more frequently observed on sugarcane, while Ms15 was almost exclusively collected in colonies on wild sorghum. Laboratory transfer experiments demonstrated the existence of fitness trade-offs. An Ms11 isofemale lineage performed better on sugarcane than on sorghum, whereas an Ms15 lineage developed very poorly on sugarcane, and two Ms16 lineages showed no significant difference in performances between both hosts. Both field and laboratory results support the existence of host plant specialization in M. sacchari in Reunion Island, despite low genetic differentiation. This study illustrates the ability of asexual aphid lineages to rapidly undergo adaptive changes including shifting from one host plant to another.

On Reunion Island successful biological control of the sugarcane white grub Hoplochelus marginalis Fairmaire (Coleoptera: Melolonthidae) has been conducted for decades with strains from the entomopathogenic fungal genus Beauveria (Ascomycota: Hypocreales). A study based on morphological characters combined with a multisequence phylogenetic analysis of genes that encode the translation elongation factor 1-alpha (TEF1), RNA polymerase II largest subunit (RPB1), RNA polymerase II second largest subunit (RPB2) and the Bloc nuc intergenic region was carried out on Beauveria strains isolated on Reunion and Madagascar from H. marginalis. This study revealed that these strains, previously identified as Beauveria brongniartii, did not match that species and are closely related to but still distinct from B. malawiensis strains. Therefore we describe the Reunion Island fungus as the new species B. hoplocheli., (© 2015 by The Mycological Society of America.)

The work consisted of two experiments, i.e. Experiment 1 was conducted under controlled environments where sugarcane plants were used as hosts. This investigation aimed to monitor the occurrence of the Sugarcane White Leaf disease and the abundance of Leafhopper insect vectors and also the work aimed to provide useful information in understanding some aspects on epidemiology of the Sugarcane White Leaf disease. A Completely Randomized Design with three replications was used to justify growth and development of Leafhopper insects as affected by different temperatures: 20 (T1), 25 (T2), 30 (T3) and 35 degrees C (T4). Experiment 2 was carried out to determine the numbers of Leafhopper insects with the use of light traps in the sugarcane Field 1 (ratoon plants), Field 2 (newly planted), Field 3 (newly planted) and Field 4 (ratoon plants). The results of Experiment 1 showed that growth and development of Leafhopper insects were highly affected by temperatures i.e. the higher the environmental temperature the faster the growth and development of the insects to reach its full adulthood. At 20 degrees C, Leafhopper insects took 12 days to lay eggs whereas at 25 degrees C the insects took only 6 days. Male reached its adulthood approximately 9 days earlier than female when cultured at 25 degrees C and became approximately one week at 30 degrees C or higher. The results of Experiment 2 showed that the majority of Leafhopper insects were found within the months of June and July for both newly planted and ratoon crops. A small amount was found in May and August with an exceptional case of Field 4 where the highest number of Leafhopper insects was found in April followed by June and July. For Sugarcane White Leaf disease, the disease was found in all months of the year except February for Fields 2 and 3. Newly planted sugarcane plants attained much smaller percentages of disease than those of the ratoon plants.

Sugarcane is a widely cultivated plant that serves primarily as a source of sugar and ethanol. Its annual yield can be significantly reduced by the action of several insect pests including the sugarcane giant borer (Telchin licus licus), a lepidopteran that presents a long life cycle and which efforts to control it using pesticides have been inefficient. Although its economical relevance, only a few DNA sequences are available for this species in the GenBank. Pyrosequencing technology was used to investigate the transcriptome of several developmental stages of the insect. To maximize transcript diversity, a pool of total RNA was extracted from whole body insects and used to construct a normalized cDNA database. Sequencing produced over 650,000 reads, which were de novo assembled to generate a reference library of 23,824 contigs. After quality score and annotation, 43% of the contigs had at least one BLAST hit against the NCBI non-redundant database, and 40% showed similarities with the lepidopteran Bombyx mori. In a further analysis, we conducted a comparison with Manduca sexta midgut sequences to identify transcripts of genes involved in digestion. Of these transcripts, many presented an expansion or depletion in gene number, compared to B. mori genome. From the sugarcane giant borer (SGB) transcriptome, a number of aminopeptidase N (APN) cDNAs were characterized based on homology to those reported as Cry toxin receptors. This is the first report that provides a large-scale EST database for the species. Transcriptome analysis will certainly be useful to identify novel developmental genes, to better understand the insect's biology and to guide the development of new strategies for insect-pest control.

The sugarcane borer moth, Diatraea saccharalis, is widespread throughout the Western Hemisphere, and is considered an introduced species in the southern United States. Although this moth has a wide distribution and is a pest of many crop plants including sugarcane, corn, sorghum and rice, it is considered one species. The objective was to investigate whether more than one introduction of D. saccharalis had occurred in the southern United States and whether any cryptic species were present. We field collected D. saccharalis in Texas, Louisiana and Florida in the southern United States. Two molecular markers, AFLPs and mitochondrial COI, were used to examine genetic variation among these regional populations and to compare the sequences with those available in GenBank and BOLD. We found geographic population structure in the southern United States which suggests two introductions and the presence of a previously unknown cryptic species. Management of D. saccharalis would likely benefit from further investigation of population genetics throughout the range of this species.

The enzymatic activity of leucine aminopeptidase (EC 3.4.11.1) from the intestinal tract of sugarcane giant borer (Telchin licus licus) was assayed by using a simple and sensitive spectrophotometric assay that uses L-leucyl-2- naphthylamide as substrate. In this assay, L-leucyl-2-naphthylamide is hydrolyzed to produce 2-naphthylamine and Lleucine. The product 2-naphthylamine reacts with Fast Black K and can be monitored using a continuous spectrophotometric measurement at 590 nm. The data on the kinetic parameters indicates that the Km for the L-leucyl-2- naphthylamide at pH 7.0 was found to be lower than those found for other LAP substrates. The Km and Vmax for the LAP were determined to be 84.03 µM and 357.14 enzymatic units mg(-1), respectively. A noticeable difference of LAP activity between the two insect orders tested was observed. This method could be used to screen for natural LAP inhibitors.

We present polymorphic microsatellite markers isolated for genetic studies of the sugarcane borer, Diatraea saccharalis (Fabricius). We isolated 16 microsatellite loci through an enriched genomic library protocol. After characterization, 12 markers showed polymorphic information expressed in the observed number of alleles (ranging from 2 to 7; 5 on average) and in the polymorphism information content (ranging from 0.292 to 0.771; 0.535 on average). These markers can be used in further studies to understand the basic ecological characteristics of the sugarcane borer, e.g., dispersion patterns and population genetic differentiation, associated with distinct geographic scales and host plants.

Root lesion nematodes (RLNs, Pratylenchus species) are a group of economically important migratory endoparasitic plant pathogens that attack host roots of major crops such as wheat and sugarcane, and can reduce crop yields by 7-15%. Pratylenchus thornei and Pratylenchus zeae were treated with double stranded RNA (dsRNA) to study gene silencing, (RNA interference, RNAi), as a potential strategy for their control. Mixed stages of nematodes of both species ingested dsRNA when incubated in a basic soaking solution in the presence of the neurostimulant octopamine. Incubation for up to 16 h in soaking solutions containing 10-50 mM octopamine, 0.1-1.0 mg/mL FITC, and 0.5-6 mM spermidine did not affect vitality. Spermidine phosphate salt hexahydrate rather than spermidine or spermidine trihydrochloride increased uptake of FITC by nematodes, and this resulted in more effective gene silencing. Silencing pat-10 and unc-87 genes of P. thornei and P. zeae resulted in paralysis and uncoordinated movements in both species, although to a higher degree in P. thornei. There was also a greater reduction in transcript of both genes in P. thornei indicating that it may be more susceptible to RNAi. For P. thornei treated with dsRNA of pat-10 and unc-87 there was a significant reduction (77-81%) in nematode reproduction on carrot mini discs over a 5 week period. The results show that RLNs are clearly amenable to gene silencing, and that in planta delivery of dsRNA to target genes in these nematodes should confer host resistance. Moreover, for the two genes, dsRNA derived from either nematode species silenced the corresponding gene in both species. This implies cross-species control of nematodes via RNAi is possible., (Copyright © 2012 Elsevier Inc. All rights reserved.)

In sugarcane fields, colonization of the stalk by opportunistic fungi usually occurs after the caterpillar Diatraea saccharalis attacks the sugarcane plant. Plants respond to insect attack by inducing and accumulating a large set of defense proteins. Two homologues of a barley wound-inducible protein (BARWIN), sugarcane wound-inducible proteins SUGARWIN1 and SUGARWIN2, have been identified in sugarcane by an in silico analysis. Antifungal properties have been described for a number of BARWIN homologues. We report that a SUGARWIN::green fluorescent protein fusion protein is located in the endoplasmic reticulum and in the extracellular space of sugarcane plants. The induction of sugarwin transcripts occurs in response to mechanical wounding, D. saccharalis damage, and methyl jasmonate treatment. The accumulation of transcripts is late induced and is restricted to the site of the wound. Although the transcripts of sugarwin genes were strongly increased following insect attack, the protein itself did not show any effect on insect development; rather, it altered fungal morphology, leading to the apoptosis of the germlings. These results suggest that, in the course of evolution, sugarwin-encoding genes were recruited by sugarcane due to their antipathogenic activity. We rationalize that sugarcane is able to induce sugarwin gene expression in response to D. saccharalis feeding as a concerted plant response to the anticipated invasion by the fungi that typically penetrate the plant stalk after insect damage.

To improve transgene expression level, we synthesized a truncated insecticidal gene m-cry1Ac by increasing its GC content from 37.4 to 54.8%, based on the codon usage pattern of sugarcane genes, and transferred it into two sugarcane cultivars (ROC16 and YT79-177) by microprojectile bombardment. The integration sites and expression pattern of the transgene were determined, respectively, by Southern, northern and western blot analyses. The transgenic sugarcane lines produced up to 50 ng Cry1Ac protein per mg soluble proteins, which was about fivefold higher than that produced by the partially modified s-cry1Ac (GC% = 47.5%). In greenhouse plant assay, about 62% of the transgenic lines exhibited excellent resistance to heavy infestation by stem borers. In field trials, the m-cry1Ac transgenic sugarcane lines expressing high levels of Cry1Ac were immune from insect attack. In contrast, expression of s-cry1Ac in transgenic sugarcane plants resulted in moderately decreased damages in internodes (0.4-1.7%) and stalks (13.3-26.7%) in comparison with the untransformed sugarcane controls, which showed about 4 and 26-40% damaged internodes and stalks, respectively. Significantly, these transgenic sugarcane lines with high levels of insect resistance showed similar agronomic and industrial traits as untransformed control plants. Taken together, the findings from this study indicate a promising potential of engineering an insect-resistant gene to tailor its protein expression levels in transgenic sugarcane to combat insect infestations.

Changes in the agroecosystem with the increase of green cane harvesting in Brazil affected the insect populations associated to this crop, and secondary pests like the spittlebug Mahanarva fimbriolata Stål, became much more important. Many studies have demonstrated the active role played by silicon in plant defense against herbivory. The objective of this study was to evaluate the effects of silicon applications on the biology of the spittlebug reared on two resistant (SP79-1011 and SP80-1816) and one susceptible (SP81-3250) sugarcane cultivars. Sugarcane plants were grown under greenhouse conditions and submitted to different treatments: with and without silicon fertilizer in two different soil type (sandy and clay soil). The newly hatched nymphs were transferred to sugarcane roots and placed into boxes with lids, to keep a moistened and dark environment favoring their growth and maintenance of the root system, providing food access to the developing nymphs. After emergence, adult males and females were placed in cages for mating and oviposition. The silicon absorbed and accumulated in the plant caused an increase in nymphal mortality, and depending on the sugarcane cultivar tested this element also provided an increase in the duration of the nymphal stage and a decrease in the longevity of males and females. 'SP79-1011' presented the highest silicon content in leaves, and M. fimbriolata had the highest nymph mortality and the shortest female longevity. The pre-oviposition period, fecundity and egg viability were not affected by the silicon content in plants or the cultivar used.

Sugarcane is an important crop that has recently become subject to attacks from the weevil Sphenophorus levis, which is not efficiently controlled with chemical insecticides. This demands the development of new control devices for which digestive physiology data are needed. In the present study, ion-exchange chromatography of S. levis whole midgut homogenates, together with enzyme assays with natural and synthetic substrates and specific inhibitors, demonstrated that a cysteine proteinase is a major proteinase, trypsin is a minor one and chymotrypsin is probably negligible. Amylase, maltase and the cysteine proteinase occur in the gut contents and decrease throughout the midgut; trypsin is constant in the entire midgut, whereas a membrane-bound aminopeptidase predominates in the posterior midgut. The cysteine proteinase was purified to homogeneity through ion-exchange chromatography. The purified enzyme had a mass of 37 kDa and was able to hydrolyze Z-Phe-Arg-MCA and Z-Leu-Arg-MCA with k(cat)/K(m) values of 20.0±1.1 μM(-1)s(-1) and 30.0±0.5 μM(-1)s(-1), respectively, but not Z-Arg-Arg-MCA. The combined results suggest that protein digestion starts in the anterior midgut under the action of a cathepsin L-like proteinase and ends on the surface of posterior midgut cells. All starch digestion takes place in anterior midgut. These data will be instrumental to developing S. levis-resistant sugarcane., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Feeding by lesser cornstalk borer, Elasmopalpus lignosellus (Zeller) (Lepidoptera: Pyralidae), larvae on sugarcane (a complex hybrid of Saccharum spp.) causes leaf damage, dead hearts, and dead plants that can result in stand and yield loss. A 2-yr greenhouse experiment was conducted to examine sugarcane variety and plant age-specific feeding responses to E. lignosellus. Plants growing from single-eye setts of three varieties were exposed to a single generation of E. lignosellus larvae beginning at the three-, five-, and seven-leaf stages. Results indicated that the physical damage and resulting yield loss of plants attacked by E. lignosellus larvae were dependent on the variety and leaf stage at which they were infested. Significantly more plant damage was observed in all three varieties when infested at the three- than at the seven-leaf stage. Larvae caused significantly more plant damage and reduced yield in CP 89-2143 than in CP 78-1628. Tiller production increased in CP78-1628 and CP 88-1762 when infested at the three-leaf stage, whereas tiller production, biomass and sugar yield decreased in CP 89-2143 when infested at all leaf stages, compared with the untreated control. There was no reduction in yield when CP 78-1628 was infested at the three- or five-leaf stages. Biomass was reduced in CP 88-1762 when plants were infested at any of the leaf stages, but sugar yield was reduced only when infested at the seven-leaf stage. These results indicate that compensation in response to E. lignosellus damage was variety dependent and declined with the delay in infestation time.

Lesser cornstalk borer, Elasmopalpus lignosellus (Zeller) (Lepidoptera: Pyralidae) is an important pest of sugarcane (a complex hybrid of Saccharum spp.) in southern Florida. Cultural controls for E. lignosellus were evaluated in preparation for the potential loss of effective insecticides. Field studies conducted in 2006 compared the effects of harvest residues from green-harvested sugarcane (no preharvest burning to remove leaf matter) on E. lignosellus stalk damage and yield. Damage by E. lignosellus was significantly lower in plant cane plots that were covered with harvest residues collected from a green-harvested field before shoot emergence compared with plots without harvest residue. There were no yield differences between plots with and without harvest residues in plant or ratoon sugarcane fields in the 2006 study. The effects of three postharvest tillage levels (conventional, intermediate, and no tillage) were evaluated in preharvest burned and green-harvested fields in 2008 and 2009. Significantly less E. lignosellus damage was observed in the green- versus preharvest burned fields in both years. Intermediate and no-tillage plots had very little damage in green-harvested field. Conventional tillage plots had the greatest damage in the green-harvested field and the lowest damage in the preharvest burned field. In 2008, biomass yield was greater in the intermediate than conventional tillage in the green-harvested field, but it was greater in the conventional than in other tillage levels in the preharvest burned field. These studies demonstrated that cultural controls could greatly reduce E. lignosellus damage in sugarcane without the use of insecticides.

The sugarcane borer, Diatraea saccharalis, is a major target pest of transgenic corn expressing Bacillus thuringiensis (Bt) proteins (i.e., Cry1Ab) in South America and the mid-southern region of the United States. Evolution of insecticide resistance in such target pests is a major threat to the durability of transgenic Bt crops. Understanding the pests' resistance mechanisms will facilitate development of effective strategies for delaying or countering resistance. Alterations in expression of cadherin- and alkaline phosphatase (ALP) have been associated with Bt resistance in several species of pest insects. In this study, neither the activity nor gene regulation of ALP was associated with Cry1Ab resistance in D. saccharalis. Total ALP enzymatic activity was similar between Cry1Ab-susceptible (Cry1Ab-SS) and -resistant (Cry1Ab-RR) strains of D. saccharalis. In addition, expression levels of three ALP genes were also similar between Cry1Ab-SS and -RR, and cDNA sequences did not differ between susceptible and resistant larvae. In contrast, altered expression of a midgut cadherin (DsCAD1) was associated with the Cry1Ab resistance. Whereas cDNA sequences of DsCAD1 were identical between the two strains, the transcript abundance of DsCAD1 was significantly lower in Cry1Ab-RR. To verify the involvement of DsCAD1 in susceptibility to Cry1Ab, RNA interference (RNAi) was employed to knock-down DsCAD1 expression in the susceptible larvae. Down-regulation of DsCAD1 expression by RNAi was functionally correlated with a decrease in Cry1Ab susceptibility. These results suggest that down-regulation of DsCAD1 is associated with resistance to Cry1Ab in D. saccharalis.

The lesser cornstalk borer, Elasmopalpus lignosellus (Zeller) (Lepidoptera: Pyralidae) is an important pest of sugarcane (a complex hybrid of Saccharum spp.) in southern Florida. Reproductive and life table parameters for E. lignosellus were examined at nine constant temperatures from 13 to 36 °C with sugarcane as the larval food source. The pre- and postoviposition periods decreased with increasing temperatures and reached their minimums at 33 and 36 °C, respectively. The oviposition period was longest at 27 °C. The mean fecundity, stage-specific survival, stage-specific fecundity, intrinsic rate of increase, and finite rate of increase were greatest at 30 °C and decreased with increasing or decreasing temperature. The net reproductive rate was greatest at 27 °C. The Logan-6 model best described the relationship between temperature and intrinsic rate of increase. The generation and population doubling times were longest at 13 and shortest at 33 and 30 °C, respectively. The most favorable temperatures for E. lignosellus population growth were between 27 and 33°C. Life table parameters for E. lignosellus reared on sugarcane were greater than for the Mexican rice borer [Eoreuma loftini (Dyar) (Lepidoptera: Crambidae)] reared on an artificial diet at 30 °C. The intrinsic rates of increase for the sugarcane borer [Diatraea saccharalis (F.) (Lepidoptera: Crambidae)] reared on sugarcane or corn were the same as for E. lignosellus reared on sugarcane at 27 °C, but the net reproductive rate was four times higher for the former than the latter borer species., (© 2010 Entomological Society of America)

Sugarcane (Saccharum spp.) in Louisiana is colonized by two aphid species, the sugarcane aphid, Melanaphis sacchari (Zehntner), and the yellow sugarcane aphid, Sipha flava (Forbes) (Hemiptera: Aphididae). The main problem associated with M. sacchari is transmission of sugarcane yellow leaf virus, a casual agent of yellow leaf disease whose absence has been added to certification standards for micropropagated sugarcane in Louisiana. Greenhouse studies were conducted to categorize dominant commercial sugarcane cultivars for their ability to tolerate aphid injury and to express antixenotic or antibiotic effects on both aphid species. Antixenosis tests showed no preference among cultivars by either aphid species. Loss of chlorophyll content in tolerance tests also did not show differences among cultivars for both aphid species. However, antibiosis tests revealed that life history parameters such as the duration of the reproductive period and fecundity of both aphid species were negatively affected on 'HoCP 91-555' compared with 'L 97-128'. Estimation of demographic statistics indicated that both aphid species exhibited a significantly lower intrinsic rate of increase (1.8-2.8-fold) and longer doubling time (1.7-3.1-fold) on HoCP 91-555 relative to L 97-128. From these tests, cultivars in the current study can be ranked from most to the least susceptible as L 97-128 > 'LCP 85-384' > 'HoCP 96-540' > 'Ho 95-988' > HoCP 91-555 for M. sacchari and L 97-128 > LCP 85-384 > HoCP 91-555 for S. flava. Therefore, antibiosis is an important category of resistance in sugarcane to both aphid species, and HoCP 91-555 might provide useful germplasm for developing aphid resistant cultivars.

The occurrence of Eldana saccharina (Lepidoptera: Pyralidae) was monitored in grids represented by plots in 12 nematicide trials in South African sugarcane fields. The trials encompassed a total of eight plant cane crops and 22 ratoon crops and were situated within commercial cane fields. Several measurements were made to characterize the damage caused by E. saccharina. These included the number of internodes per stalk, the percentage of internodes damaged and the percentage of stalks damaged. The mapping of E. saccharina infestation in plant crops of sugarcane showed that the borders of the trials were as infested as the centre, indicating invasion from outside the field plus internal spread within the field. Ratoon crops were less infested than plant crops. This could be explained by a shorter ratoon crop cycle and by the fields having areas that were more suitable for the borer than elsewhere. The location of these preferred areas could be predicted from one ratoon crop to the next but was not related to the distribution of the borer in the plant crop. This situation was thought to explain the apparent stabilization of E. saccharina infestation in ratoon cane. Because the borer was found at harvest only in stalks with more than 14 to 16 internodes, it appeared that the oldest shoots, or the shoots with the greatest growth potential, attracted the insect, possibly due to their higher nitrogen content, which would stimulate growth. All the trials were on sandy soil, and crop loss from nematodes was greater than that caused by E. saccharina.