Rice Strains Research Articles

PCR molecular identification of the fall armyworm in the Chontalpa region, Tabasco, Mexico

O Objective: To perform the first molecular characterization of the barcoding COI region of the S. frugiperda found in Tabasco and to test whether it can reliably identify the strain detected in the said state. Design/Methodology/Approach: We collected S. frugiperda specimens using four pheromone traps placed in corn plots. Subsequently, we implemented DNA extraction, PCR with LCO-L/HCO-L primers, and the sequencing of six individuals captured in the traps. Results: After aligning the sequences, we developed a phylogenetic tree, determining that S. frugiperda belongs to the rice strain (RS). Implications: Identifying the S. frugiperda strain is necessary for management purposes, since different strains may require different control methods. Findings/Conclusions: Our study revealed that the isolates from Chontalpa, Tabasco, Mexico, match the RS fall armyworm reported in other latitudes. Consequently, this is the first report to identify this strain of S. frugiperda in Tabasco. Our findings are relevant because this species can potentially become a pest in sugarcane- and rice-growing areas of the state.

Identification of Spodoptera frugiperda (Lepidoptera: Noctuidae) and its two host strains in China by PCR-RFLP.

The fall armyworm (FAW) Spodoptera frugiperda was first found in China in 2018. In other countries, FAW has evolved corn and rice strain biotypes. It is not possible to identify these strains based on morphology. In addition, FAW is very similar in appearance to several other common pests. These situations bring great challenges to the population management of FAW. In this study, we developed a rapid identification method based on PCR-RFLP to distinguish the two FAW strains and the FAW from other lepidopteran pests. A 697 bp mitochondrial cytochrome c oxidase I (COI) was cloned and sequenced from FAW, Spodoptera litura, Spodoptera exigua, and Mythimna separata. The COI fragments of these species revealed unique digestion patterns created by three enzymes (Tail, AlWN I, and BstY II). Thus, these four species can be distinguished from each other. The enzyme Ban I recognized a unique SNP site on a 638 bp triosephosphate isomerase (Tpi) fragment of the corn strain FAW. The Tpi fragment of the corn strain was cut into two bands. However, the rice strain could not be digested. Using this method, all 28 FAW samples collected from different host plants and locations in China were identified as the corn strain. This suggests that the rice strain has not yet invaded China. This method allows discrimination of FAW from other Lepidopteran pests and distinguishes the two FAW host strains.

Population genomics of fall armyworm by genotyping-by-sequencing: Implications for pest management.

The fall armyworm (FAW), Spodoptera frugiperda, is a significant pest of many crops in the world and it is native to the Americas, where the species has shown the ability to rapidly evolve resistance to insecticides and transgenic plants. Despite the importance of this species, there is a gap in the knowledge regarding the genetic structure of FAW in South America. Here, we examined the genetic diversity of FAW populations across a wide agricultural area of Brazil and Argentina using a Genotyping-by-Sequencing (GBS) approach. We also characterized samples by their host strain based on mitochondrial and Z-linked genetic markers. The GBS methodology enabled us to discover 3309 SNPs, including neutral and outlier markers. Data showed significant genetic structure between Brazil and Argentina populations, and also among the Argentinian ecoregions. Populations inside Brazil showed little genetic differentiation indicating high gene flow among locations and confirming that structure is related to the presence of corn and rice strains. Outlier analysis indicated 456 loci putatively under selection, including genes possibly related to resistance evolution. This study provides clarification of the population genetic structure of FAW in South America and highlights the importance of genomic research to understand the risks of spread of resistance genes.

Biology of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) on Rice and Different Corn Varieties

Rice and corn are one of the most important crops in the Philippines. Several insect pests contribute to the losses and low yield of these crops. The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), was recently reported to cause economic damage to corn. Also, this insect consists of two genetically differentiated strains namely, the corn strain and the rice strain. This study was conducted in a laboratory conditions to evaluate the biological parameters of FAW on rice and corn varieties. Newly hatched larvae were fed such as the open pollinated variety (OPV), traditional variety, Macho F1 (hybrid), NK 6410 (genetically modified organism), and rice variety, RC 226. Duration of larva, prepupal, pupal, and larva-adult period, pre-oviposition, oviposition, and post-oviposition period, number of egg mass, fecundity, sex ratio, and survivability of larva to adult of different crops were evaluated. The results showed that the duration of development from egg to adult in Macho F1 and NK 6410 with an average duration of 24.58 and 24.66 d was significantly shorter than OPV, traditional variety, and rice variety, RC 226. The number of eggs laid by females reared on Macho F1, NK 6410, and OPV, with a mean of 1125.60, 1176.80, and 1049.40, respectively, were significantly greater than the traditional variety, tinigib, and rice RC 226. Different corn varieties and rice did not affect the male and female ratio, longevity, and survivability. The results of this study will provide insights into formulating management strategies for fall armyworm.

A CRISPR/Cas12a-Mediated Sensitive DNA Detection System for Gene-Edited Rice.

In recent years, genome editing technology represented byclustered regularly interspaced short palindromic repeat/CRISPR-associated nuclease 9 （CRISPR/Cas9） has been developed and applied in transgenic research and development, and transgenic products have been developed for a variety of applications. Gene editing products, unlike traditional genetically modified crops, which are generally obtained by target gene deletion, insertion, base mutation, etc., may not differ significantly at the gene level from conventional crops, which increases the complexity of testing. We established a specific and sensitive CRISPR/Cas12a-mediated gene editing system to detect target fragments in a variety of transgenic rice lines and commercial rice-based processing products. In this study, the CRISPR/Cas12a visible detection system was optimized for the visualization of nucleic acid detection in gene-edited rice. The fluorescence signals were detected by both gel electrophoresis and fluorescence-based methods. The detection limit of the CRISPR/Cas12a detection system established in this study was more precise, especiallyfor low-concentration samples. In addition to achieving single-base detection in gene-edited rice, we showed that different base mutations in the target sequence have different detection efficiencies by sitewise variant compact analysis. The CRISPR/Cas12a system was verified via a common transgenic rice strain and commercial rice sources. The results proved that the detection method could not only be tested in samples with multiple mutation types but could also effectively detect target fragments in commercial rice products. We have developed a set of efficient detection methods with CRISPR/Cas12a for gene-edited rice detection to provide a new technical basis for rapid field detection of gene-edited rice. The CRISPR/Cas12a-mediated visual detection method used to detect gene-edited rice was evaluated for its specificity, sensitivity, and robustness.

Comparative transcriptome-wide identification and differential expression of genes and lncRNAs in rice near-isogenic line (KW-Bph36-NIL) in response to BPH feeding.

Brown planthopper (BPH) is the most devastating pest of rice in Asia, causing substantial yield losses and has become a challenging task to be controlled under field conditions. Although extensive measures have been taken over the past decades, which resulted in the evolution of new resistant BPH strains. Therefore, besides other possible approaches, equipping host plants with resistant genes is the most effective and environment-friendly technique for BPH control. Here, we systematically analyzed transcriptome changes in the susceptible rice variety Kangwenqingzhan (KW) and the resistant near-isogenic line (NIL) KW-Bph36-NIL, through RNA-seq, depicting the differential expression profiles of mRNAs and long non-coding RNAs (lncRNAs) in rice before and after BPH feeding. We observed a proportion of genes (1.48%) and (2.74%) were altered in KW and NIL, respectively, indicating different responses of rice strains against BPH feeding. Nevertheless, we characterized 384 differentially expressed long non-coding RNAs (DELs) that can be impacted by the two strains by alternatively changing the expression patterns of the respective coding genes, suggesting their certain involvement in response to BPH feeding. In BPH invasion, KW and NIL responded differently by modifying the synthesis, storage, and transformation of intracellular substances, adjusting the nutrient accumulation and utilization inside and outside the cells. In addition, NIL expressed stronger resistance by acutely up-regulating genes and other transcription factors related to stress resistance and plant immunity. Altogether, our study elaborates valuable insights into the genome-wide DEGs and DELs expression profiles of rice under BPH invasion by high throughput sequencing and further suggests that NILs can be utilized in BPH resistance breeding programs in developing high-resistance rice lines.

Precise Authenticity of Quinoa, Coix Seed, Wild Rice and Chickpea Components Using Optimized TaqMan Real-Time PCR.

Functional food such as, quinoa, coix seed, wild rice and chickpea have experienced rapidly increasing demand globally and exhibit high economic values. Nevertheless, a method for rapid yet accurate detection of these source components is absent, making it difficult to identify commercially available food with labels indicating the presence of relevant components. In this study, we constructed a real-time quantitative polymerase chain reaction (qPCR) method for rapid detection of quinoa, coix seed, wild rice and chickpea in food to identify the authenticity of such food. Specific primers and probes were designed with 2S albumin genes of quinoa, SAD genes of coix seed, ITS genes of wild rice and CIA-2 genes of chickpea as the target genes. The qPCR method could specifically identify the four wild rice strains, yielding, LODs of 0.96, 1.14, 1.04 and 0.97 pg/µL quinoa, coix seed, wild rice and chickpea source components, respectively. Particularly, the method allowed the identification of the target component with content below 0.01%. A total of 24 commercially available food samples of different types were detected by using the method and the results indicate that the developed method is applicable to the detection of different food matrices, as well as authenticity verification in deeply processed food.

Genetic analyses and detection of point mutations in the acetylcholinesterase-1 gene associated with organophosphate insecticide resistance in fall armyworm (Spodoptera frugiperda) populations from Uganda

BackgroundThe fall armyworm (FAW), Spodoptera frugiperda; J.E. Smith (Lepidoptera: Noctuidae), is now an economically important pest that causes huge losses to maize productivity in sub-Saharan Africa. Variations in sub-population genetics and the processes of rapid adaptation underpinning the invasion remain unclear. For this, the genetic identity and diversity of FAW populations in Uganda were revealed by sequencing 87 samples (collected across the country). Based on the partial mitochondrial cytochrome oxidase I (COI) gene polymorphisms, we further examined the mitochondrial haplotype configuration and compared the FAW in Uganda with sequences from other parts of the world. The molecular target for organophosphate and carbamate resistance, acetylcholinesterase, was also investigated.ResultsAnalysis of the partial COI gene sequences showed the presence of both rice (predominant) and corn strain haplotypes, with a haplotype diversity of 0.382. Based on the COI marker, pairwise difference distribution analyses, and neutrality tests, showed that the FAW populations in Uganda and the rest of Africa are evolving neutrally, but those in America and Asia are undergoing expansion. Our findings support observations that invasive FAW populations throughout the rest of Africa and Asia share a common origin. Sequencing of the S. frugiperda ace-1 gene revealed four amino acid substitutions, two of which (A201S and F290V) were previously shown to confer organophosphate resistance in both S. frugiperda and several other insect species. The other two previously reported new variations in positions g-396 and g-768, are presumed to be related to the development of insecticide resistance.ConclusionsThis research has increased our knowledge of the genetics of FAW in Uganda, which is critical for pest surveillance and the detection of resistance. However, due to the low gene polymorphism of COI, more evolutionary studies incorporating the Spodoptera frugiperda whole-genome sequence are required to precisely understand the FAW population dynamics, introduction paths, origin, and subsequent spread.

DEVELOPMENT OF NEW RICE STRAIN WITH IMPROVED SINK SIZE AND SOURCE CAPACITY USING A MULTI-PARENT ADVANCED GENERATION INTERCROSS (MAGIC) APPROACH

A new rice strain that grows six feet tall with four times higher potential yield than the conventionally bred variety, JP5, was developed using the MAGIC approach. This new rice type underwent analysis on sink size and source capacity traits, including superior and inferior spikelets, vascular bundles of panicle neck and stem internode, tillering pattern, grain filling pattern, yield, and other morphological attributes. Results indicated that the new strain had more vascular bundles of the stem (42) and panicle neck (35), primary rachis branches of panicle (16.1), superior spikelets, and greater grain weight than the conventionally bred variety, JP5. The panicle measured 45 cm long, with fertile grains of 500 per panicle and a stem diameter of 1.2 cm. During the grain filling duration, the spikelets of this strain and superior spikelets of JP5 gained maximum weight earlier than the inferior spikelets of JP5. Six feet tall plants of this new strain with long and heavy panicles had greater stem wall thickness. There occurred a positive and significant correlation (0.97) between yield and small vascular bundles of the panicle neck, lumen diameter (0.98), leaf length (0.99), leaf width (0.99), flag leaf length (0.99*), flag leaf width (0.97<), panicle length (0.97), fertile grains per panicle (0.98), and plant height (0.97*). The study noted that improving sink size, source capacity, and transportation of assimilation contributed positively toward yield. This novel strategy for grain yield enhancement in rice proved beneficial for other cereals to get significant breakthroughs in their production for ensuring food security.

Combined Effects of BEIIb and SSIIa Alleles on Amylose Contents, Starch Fine Structures and Physicochemical Properties of Indica Rice

Starch branching enzyme IIb (BEIIb) and soluble starch synthase IIa (SSIIa) play important roles in starch biosynthesis in cereals. Deficiency in the BEIIb gene produces the amylose extender (ae) mutant rice strain with increased amylose content (AC) and changes in the amylopectin structure. The SSIIa gene is responsible for the genetic control of gelatinization temperature (GT). The combined effects of BEIIb and SSIIa alleles on the AC, fine structures, and physicochemical properties of starches from 12 rice accessions including 10 recombinant inbred lines (RIL) and their two parents were examined in this study. Under the active BEIIb background, starches with the SSIIa-GC allele showed a higher GT than those with the SSIIa-TT allele, resulting from a lower proportion of A chain and a larger proportion of B1 chains in the amylopectin of SSIIa-GC. However, starch with the BEIIb mutant allele (be2b) in combination with any SSIIa genotype displayed more amylose long chains, higher amylose content, B2 and B3 chains, and molecular order, but smaller relative crystallinity and proportion of amylopectin A and B1 chains than those with BEIIb, leading to a higher GT and lower paste viscosities. These results suggest that BEIIb is more important in determining the structural and physicochemical properties than SSIIa. These results provide additional insights into the structure-function relationship in indica rice rather than that in japonica rice and are useful for breeding rice with high amylose content and high resistant starch.

Active microbial arsenic methylation in saline-alkaline paddy soil

Seawater rice has been cultivated to ensure food security. The salt-tolerant rice strains are resistant to saline and alkali but may be vulnerable to elevated arsenic (As) near coastal regions. Herein, the saline-alkaline paddy soil was incubated with natural irrigation river for three months to explore the mobility and transformation of As. The incubation results showed that 65 ± 1.2 % solid-bound As(V) was reduced to As(III) within two weeks with the release of As(III) to porewater. The dissolved As(III) was methylated after two weeks, resulting in dimethyl arsenate (DMA) as the dominant As species (87 %–100 %). The elevated As methylation was attributed to the most abundant arsenite methyltransferase gene (arsM) (4.1–10.4 × 107/g dry soil), over three orders of magnitude higher than As redox-related genes. The analysis of arsM operational taxonomic units (OTUs) suggested the highest sequence similarity to Proteobacteria (25.7–39.5 %), Actinobacteria (24.9–30.5 %), Gemmatimonadetes (7.5–11.9 %), Basidiomycota (5.1–12.5 %), and Chloroflexi (4.1–8.7 %). Specifically, Chloroflexi and Actinobacteria are salt-tolerant bacteria, probably responsible for As methylation. The As in grain was within a safe regulatory level, and the dominance of methylated As in porewater did not enhance its accumulation in rice grains.

Application of data integration for rice bacterial strain selection by combining their osmotic stress response and plant growth-promoting traits.

Agricultural application of plant-beneficial bacteria to improve crop yield and alleviate the stress caused by environmental conditions, pests, and pathogens is gaining popularity. However, before using these bacterial strains in plant experiments, their environmental stress responses and plant health improvement potential should be examined. In this study, we explored the applicability of three unsupervised machine learning-based data integration methods, including principal component analysis (PCA) of concatenated data, multiple co-inertia analysis (MCIA), and multiple kernel learning (MKL), to select osmotic stress-tolerant plant growth-promoting (PGP) bacterial strains isolated from the rice phyllosphere. The studied datasets consisted of direct and indirect PGP activity measurements and osmotic stress responses of eight bacterial strains previously isolated from the phyllosphere of drought-tolerant rice cultivar. The production of phytohormones, such as indole-acetic acid (IAA), gibberellic acid (GA), abscisic acid (ABA), and cytokinin, were used as direct PGP traits, whereas the production of hydrogen cyanide and siderophore and antagonistic activity against the foliar pathogens Pyricularia oryzae and Helminthosporium oryzae were evaluated as measures of indirect PGP activity. The strains were subjected to a range of osmotic stress levels by adding PEG 6000 (0, 11, 21, and 32.6%) to their growth medium. The results of the osmotic stress response experiments showed that all bacterial strains accumulated endogenous proline and glycine betaine (GB) and exhibited an increase in growth, when osmotic stress levels were increased to a specific degree, while the production of IAA and GA considerably decreased. The three applied data integration methods did not provide a similar grouping of the strains. Especially deviant was the ordination of microbial strains based on the PCA of concatenated data. However, all three data integration methods indicated that the strains Bacillus altitudinis PB46 and B. megaterium PB50 shared high similarity in PGP traits and osmotic stress response. Overall, our results indicate that data integration methods complement the single-table data analysis approach and improve the selection process for PGP microbial strains.

Characterization of lodging variation of weedy rice.

Weedy rice (Oryza spp.), one of the most notorious weeds of cultivated rice, evades eradication through stem lodging and seed shattering. Many studies have focused on seed shattering, whereas variations in lodging have received less attention and the underlying mechanisms that cause the differences in lodging between weedy and cultivated rice have not been studied in detail. Here, we compared lodging variation among diverse Chinese weedy rice strains and between weedy rice and co-occurring cultivated rice. The chemical composition of basal stems was determined, and transcriptome and methylome sequencing were used to assess the variation in expression of lodging-related genes. The results showed that the degree of lodging varied between indica-derived weed strains with high lodging levels, which occurred predominantly in southern China, and japonica-derived strains with lower lodging levels, which were found primarily in the north. The more lodging-prone indica weedy rice had a smaller bending stress and lower lignin content than non-lodging accessions. In comparison to co-occurring cultivated rice, there was a lower ratio of cellulose to lignin content in the lodging-prone weedy rice. Variation in DNA methylation of lignin synthesis-related OsSWN1, OsMYBX9, OsPAL1, and Os4CL3 mediated the differences in their expression levels and affected the ratio of cellulose to lignin content. Taken together, our results show that DNA methylation in lignin-related genes regulates variations in stem strength and lodging in weedy rice, and between weed strains and co-occurring cultivated rice.

Host-plant adaptation as a driver of incipient speciation in the fall armyworm (Spodoptera frugiperda)

BackgroundDivergent selection on host-plants is one of the main evolutionary forces driving ecological speciation in phytophagous insects. The ecological speciation might be challenging in the presence of gene flow and assortative mating because the direction of divergence is not necessarily the same between ecological selection (through host-plant adaptation) and assortative mating. The fall armyworm (FAW), a major lepidopteran pest species, is composed of two sympatric strains, corn and rice strains, named after two of their preferred host-plants. These two strains have been hypothesized to undergo incipient speciation, based on (i) several lines of evidence encompassing both pre- and post-zygotic reproductive isolation, and (ii) the presence of a substantial level of genetic differentiation. Even though the status of these two strains has been established a long time ago, it is still yet to be found whether these two strains indeed exhibit a marked level of genetic differentiation from a large number of genomic loci. Here, we analyzed whole genome sequences from 56 FAW individuals either collected from pasture grasses (a part of the favored host range of the rice strain) or corn to assess the role of host-plant adaptation in incipient speciation.ResultsPrincipal component analysis of whole genome data shows that the pattern of divergence in the fall armyworm is predominantly explained by the genetic differentiation associated with host-plants. The level of genetic differentiation between corn and rice strains is particularly marked in the Z chromosome. We identified one autosomal locus and two Z chromosome loci targeted by selective sweeps specific to rice strain and corn strain, respectively. The autosomal locus has both increased DXY and FST while the Z chromosome loci had decreased DXY and increased FST.ConclusionThese results show that the FAW population structure is dominated by the genetic differentiation between corn and rice strains. This differentiation involves divergent selection targeting at least three loci, which include a locus potentially causing reproductive isolation. Taken together, these results suggest the evolutionary scenario that host-plant speciation is a driver of incipient speciation in the fall armyworm.

Seed germination responses to temperature and water availability in weedy rice.

Weedy rice is one of the main agricultural weeds infesting transplanted and direct-seeded rice cultures across Japan. However, the environmental factors that facilitate its proliferation are largely unknown. This study investigated the effects of temperature and water availability on seed germination of Japanese weedy and cultivated rice strains differing in degree of seed dormancy and after-ripening. This information could help predict weedy rice infestation and design efficacious control measures. The germination responses of weedy rice to temperature and water availability varied among strains and after-ripening duration. The germination rate varied inversely with the degree of dormancy, increased with water availability, and fluctuated between low and high temperatures. The shallow-dormant tropical japonica-derived strawhull weedy rice (TRJ) germinated at 10°C unlike the temperate japonica-derived weedy rice strains, namely, the intermediate-dormant blackhull (BH) and the deep-dormant strawhull (SH). Seeds of all plant strains germinated below the permanent wilting point of clay loam soil, but the germination rates were higher for TRJ and BH than for cultivated rice. SH germination was more synchronous with rice than the other weedy rice strains. This study has confirmed the overall robustness of germination cold tolerance at 10°C in TRJ, high germinability at low-water availability and high temperature in TRJ and BH, and relatively more synchronous germination between SH and cultivated rice in seeds grown in different years. These observed characteristics are considered advantageous to the coexistence of weedy rice with cultivated rice. © 2022 Society of Chemical Industry.

Biological Control of Fall Armyworm, Spodoptera frugiperda

The fall armyworm (FAW), Spodoptera frugiperda, is one of the most important invasive pests worldwide, resulting in considerable losses in host crops. FAW comprises two genetic strains, such as the “rice strain”, which prefers rice and other grass species, and the “maize strain”, which feeds upon maize and sorghum. Potential control measures are generally more applicable to the farmers who lack financial assets to buy chemical insecticides or costly pure seeds. The adverse effects of pesticides on the ecosystem and human’s health and the development of resistance to insect pests have exaggerated efforts to find an alternative strategy that is cost-effective, low-risk and target-specific. Therefore, biological control is widely considered as one of the most important options for insect pest management. This comprehensive review amasses the information on biological control in all phases of their development, including predators, parasitoids, entomopathogenic fungi, viruses, nematodes, bacteria, and biopesticides, with a special focus on their effectiveness against FAW. The findings regarding biological control are briefly discussed in light of improving management programs of the invasive pest S. frugiperda.

Hybrid-derived weedy rice maintains adaptive combinations of alleles associated with seed dormancy.

Plant hybridization is a pathway for the evolution of adaptive traits. However, hybridization between adapted and nonadapted populations may affect the persistence of combinations of adaptive alleles evolved through natural selection. Seed dormancy is an adaptive trait for weedy rice because it regulates the timing of seed germination and the persistence of the soil seed bank. Hybridization between weedy and cultivated rice has been confirmed with an adaptive introgression of deep seed dormancy alleles from cultivated rice. Here, we explored the influence of hybridization on the conservation of adaptive allele combinations by evaluating natural variation and genetic structure in seed dormancy-associated genomic regions. Based on sequence variation in the genomic regions associated with seed dormancy, hybrid-derived weedy rice strains maintained most of the adaptive combinations for this trait observed in the parental weedy rice, despite equal representation of the parental weedy and cultivated rice in the whole genome sequence. Moreover, hybrid-derived weedy rice strains were more dormant than their parental weedy rice strains, and this trait was strongly influenced by the environment. This study suggests that hybridization between weedy rice (adaptive allelic combinations for seed dormancy) and cultivated rice (nonadaptive combinations) generates weedy rice strains expressing deep seed dormancy caused by genome stabilization through the removal of alleles derived from cultivated rice, in addition to the adaptive introgression of deep seed dormancy alleles derived from cultivated rice. Thus, hybridization between adapted and nonadapted populations appears to be reinforcing the trajectory towards the evolution of adaptive traits.

Comprehensive survey of transposon mPing insertion sites and transcriptome analysis for identifying candidate genes controlling high protein content of rice.

Rice is the most important crop species in the world, being staple food of more than 80% of people in Asia. About 80% of rice grain is composed of carbohydrates (starch), with its protein content as low as 7–8%. Therefore, increasing the protein content of rice offers way to create a stable protein source that contributes to improving malnutrition and health problems worldwide. We detected two rice lines harboring a significantly higher protein content (namely, HP5-7 and HP7-5) in the EG4 population. The EG4 strain of rice is a unique material in that the transposon mPing has high transpositional activity and high copy numbers under natural conditions. Other research indicated that mPing is abundant in the gene-rich euchromatic regions, suggesting that mPing amplification should create new allelic variants, novel regulatory networks, and phenotypic changes in the EG4 population. Here, we aimed to identify the candidate genes and/or mPing insertion sites causing high protein content by comprehensively identifying the mPing insertion sites and carrying out an RNA-seq-based transcriptome analysis. By utilizing the next-generation sequencing (NGS)-based methods, ca. 570 mPing insertion sites were identified per line in the EG4 population. Our results also indicated that mPing apparently has a preference for inserting itself in the region near a gene, with 38 genes in total found to contain the mPing insertion in the HP lines, of which 21 and 17 genes were specific to HP5-7 and HP7-5, respectively. Transcriptome analysis revealed that most of the genes related to protein synthesis (encoding glutelin, prolamin, and globulin) were up-regulated in HP lines relative to the control line. Interestingly, the differentially expressed gene (DEG) analysis revealed that the expression levels of many genes related to photosynthesis decreased in both HP lines; this suggests the amount of starch may have decreased, indirectly contributing to the increased protein content. The high-protein lines studied here are expected to contribute to the development of high protein-content rice by introducing valuable phenotypic traits such as high and stable yield, disease resistance, and abundant nutrients.

First report of occurrence of corn and rice strains of fall armyworm, Spodoptera frugiperda in South Sumatra, Indonesia and its damage in maize

Spodoptera frugiperda is a new invasive pest in Indonesia and its severity in maize ranges from 26.50 to 100%. However, information on the strains or genetic diversity of the S. frugiperda in Indonesia is still very limited. This research aimed to identify the genetic diversity of S. frugiperda from South Sumatra and determine its damage in maize. Surveys from January to June 2021 were carried out from the lowlands to highlands of South Sumatra. The field scouting was performed to calculate the incidence of damage and to estimate the severity caused by S. frugiperda larvae. The severity was assessed using a visual rating scale from 1 (no damage) to 5 (plant stunting and funnel damaged severely). The S. frugiperda larvae was identified based on morphological characters and molecular techniques using sequence analysis of Cytochrome c Oxidase subunit I ( COI ) gene. All larvae collected from South Sumatra showed identical morphological characteristics identified as S. frugiperda . The sequence analysis results showed that the 6 isolates of S. frugiperda shared 100% of sameness as the rice strain haplotype 1, S. frugiperda isolate from Lampung Province. The other 3 isolates of S. frugiperda shared 100% sameness as the corn strain haplotype I and IS 1 (obtained from sugarcane in Japan). All isolates have been deposited in the GenBank. This study confirmed the presence of rice and corn strains of Spodoptera frugiperda and this is the first report of the occurrence of both strains in South Sumatra. We also found that outbreaks of S. frugiperda have occured in the South Sumatra. The incidence and severity of S . frugiperda reached 100% and 65% respectively. Comprehensive further study should be performed to confirm the presence of both strains and their damage in all corn producing areas in Indonesia.

Effects of intergenerational host plant alterations on developmental parameters of Spodoptera frugiperda (Lepidoptera: Noctuidae)

• The fall armyworm (FAW) could be bred successfully on sudan grass, millet, and rye (R). • FAW’s developmental parameters showed dynamic changes by sequential alternation of host plants. • FAW was able to successfully reproduce up to five generations with non-preferred host plants. • Rye, a winter crop, might be an important hosts for migrated corn strain FAW. The fall armyworm (FAW), Spodoptera frugiperda (Smith), is known as a destructive moth pest of agricultural crops such as corn, rice, and sorghum. Recently, its damage threatens crops in Asian temperate regions due to its high migration ability. Two genetic types of FAW, a corn strain mostly found on corn and a rice strain associated with rice, have been reported. The objective of this study was to evaluate whether migrated corn-collected population of FAW could maintain its population on various seasonal plants including non-preferred Poaceae plants, with special emphasis on Jeju, Korea. Development-related parameters were compared after switching host plants including corn (C), Sudan grass (S), proso millet (M), and rye (R) when rearing FAW. Most developmental parameters showed dynamic changes according to sequential alternation of host plants with the progress of generations. When all R 0 (net reproductive rate) values were scaled by the R 0 in C to C alteration, the scaled values of other alterations were less than “1.0”, indicating less fitness than C to C alteration ( R 0 value: 0.57 for C to S, 0.49 for C to M in F3 generation; 0.82 for S to S, 0.65 for S to M, and 0.58 for S to C in F4 generation, 0.50 for M to M, 0.59 for M to R, and 0.83 for M to C in F5 generation). Although our results did not show that S. frugiperda adapted perfectively on new host plants, this pest was able to successfully reproduce up to five generations with non-preferred host plants. Results indicate that winter crops such as rye might be hosts for migrated corn-preferred populations of S . frugiperda during winter after breeding three to four cycles on other host plants before winter if weather condition is allowed for its survival.