Abnormal Leaf Research Articles

First detection of fungal diversity associated with Prunus salicina Lindl malformation syndrome

The Chinese deciduous fruit Prunus salicina Lindl is loved by people because of its distinctive flavor and significant industrial worth. This study presents the first report of abnormal leaf bud in Prunus salicina Lindl. Malformation of Prunus salicina Lindl leaf buds leads to abnormal development and growth, causing deformities and a decrease in fruit yield. Identifying the underlying causes and causative pathogens is essential for effective disease management. In this experiment, we observed the Malformation Syndrome in Prunus salicina Lindl and used the internal transcribed spacer (ITS) to compare fungal diversity between malformed and healthy leaf buds. These results show a higher level of fungal diversity in diseased leaf buds. Alternaria, Curvibasidium, Symmetrospora, Fonsecazyma, and other genera among the identified fungi are suspected to induce the malformations. Potential contributors to these malformations include Endophyte-plant_Pathogen and Plant_pathogen-Undefined_saprotroph. These findings provide valuable insights for further research on the etiology of plum bud diseases, identifying potential pathogens, and formulating control strategies.

Strategies for identification of elite Hevea brasiliensis genotypes tolerant to ALF disease from half-sib and full-sib population

Natural Rubber (NR) from Hevea brasiliensis tree is an important commercial commodity worldwide. In general, many Hevea cultivars are vulnerable to abnormal leaf fall (ALF) disease caused by Phytophthora spp., which can lead to considerable reduction in crop production. Chemical control of ALF by spraying mineral oil-based copper fungicides is the conventional approach which is harmful to the ecosystem. Development of ALF tolerant varieties of Hevea through conventional breeding is the major objectives of this study. We evaluated the primary agronomy traits and ALF tolerance level of the newly developed Hevea clones using in vitro leaf disc bioassay as well as field screening for disease incidence consecutively for three years. Variation in disease infestation was observed among genotypes. Based on field screening, the clones which showed better field tolerance against ALF than the parental clones were selected. Clone 683 was identified as one of the most desirable genotypes which had vigorous growth as well as high crop yield in addition to ALF tolerance. The study also confirms the reliability of in vitro leaf disc bioassay technique for screening against ALF across genotypes for its inherent variability. This technique is rapid and easier and can be completed within 96 hours. This approach could reduce the burden of screening larger breeding population to a manageable size to enhance the efficiency of selection in ALF disease resistance breeding programs. Selected fast growing, high yielding ALF tolerant clones, could be further evaluated in on farm trials for recommending to the growers in the traditional region of rubber growing tracts.

Insights into pathogenicity mechanisms of Phytophthora meadii, the abnormal leaf fall pathogen in rubber tree, through whole genome sequencing and analysis

Hevea brasiliensis (Willd. ex A.Juss.) Müll. Arg., also known as the Para rubber tree is the main commercial source of natural rubber. Phytophthora meadii causing abnormal leaf fall (ALF) disease is the most destructive and recurrent disease of rubber trees in India, leading to significant annual economic losses. To comprehend its pathogenic mechanisms and investigate stratagems for disease control, whole-genome sequencing and analysis of P. meadii were conducted. The P. meadii genome was sequenced using the standard Illumina Novaseq 6000 Platform. The assembled P. meadii genome is approximately 53 Mb in size, consisting of 13,171 contigs with an N50 of 7105 and a BUSCO completeness of 96 %. A total of 14,646 protein-coding genes were predicted, with an average GC content of 49.96 %. P. meadii's genome contains 750 carbohydrate-active enzymes (CAZymes), which is notably higher compared to closely related species. Five genomic regions potentially encoding enzymes for secondary metabolite production and 1877 pathogenic proteins were also identified. Our data confirm the phylogenetic positioning of P. meadii as a distinct species and indicate that it forms a closely related clade with P. citrophthora. Analyzing the P. meadii genome not only aids in understanding its genomic traits but also facilitates the identification of genes related to pathogenesis and will give a better understanding of the Hevea – Phytophthora interaction.

Influence of polyethylene microplastics on Brassica rapa: Toxicity mechanism investigation

Polyethylene microplastics (PE-MPs) have toxicity to ecological environment, including animals and plants. This study investigated the toxicity of photodegraded PE-MPs on Brassica rapa, which is a typical model plant and only have around a 30-day life cycle. It is noted that the presence of photodegraded PE-MPs inhibited Brassica rapa growth since the stem length decreased by 11.94%–51.11 % while the fresh weight and dry weight decreased by 18.56%–27.46 % and 1.90 %–6.91 % respectively, compared to the blank group. PE-MPs receiving more light radiation became more hydrophobic. This inhibited PE-MPs entering the plant body along with the process of plant absorbing water. Furthermore, when PE-MPs were located in the lower soil layers, Brassica rapa reaching them needs a longer time, hence showing lower toxicity effect than the case of PE-MPs located in the upper soil layer. The research outcomes also indicated that malondialdehyde (MDA) contents in photodegraded PE-MPs exposure group increased by 1.37%–7.28 % while the catalase activity (CAT) increased by 60.11 %. This means that PE-MPs caused oxidative stress response in plants, inducing plants to resist external stress. Transcriptomic analysis results showed that Brassica rapa, which was affected by PE-MPs, significantly up-regulated genes related to the plant-pathogen interaction pathway while the ribosome pathway genes were significantly down-regulated. This led to a decrease in growth rate and a decrease in the homeostatic level of the ribosomal subunit and hence resulting in abnormal leaf vein development. These conclusions indicated the toxic effect and damage mechanism of photodegraded PE-MPs on Brassica rapa. The novelty of this study was to use both univariate analysis and transcriptomic analysis to investigate how photodegraded PE-MPs exert toxicity on Brassica rapa. The results can provide a theoretical basis for revealing the influence of MPs on plant growth.

GHCU, a Molecular Chaperone, Regulates Leaf Curling by Modulating the Distribution of KNGH1 in Cotton.

Leaf shape is considered to be one of the most significant agronomic traits in crop breeding. However, the molecular basis underlying leaf morphogenesis in cotton is still largely unknown. In this study, through genetic mapping and molecular investigation using a natural cotton mutant cu with leaves curling upward, the causal gene GHCU is successfully identified as the key regulator of leaf flattening. Knockout of GHCU or its homolog in cotton and tobacco using CRISPR results in abnormal leaf shape. It is further discovered that GHCU facilitates the transport of the HD protein KNOTTED1-like (KNGH1) from the adaxial to the abaxial domain. Loss of GHCU function restricts KNGH1 to the adaxial epidermal region, leading to lower auxin response levels in the adaxial boundary compared to the abaxial. This spatial asymmetry in auxin distribution produces the upward-curled leaf phenotype of the cu mutant. By analysis of single-cell RNA sequencing and spatiotemporal transcriptomic data, auxin biosynthesis genes are confirmed to be expressed asymmetrically in the adaxial-abaxial epidermal cells. Overall, these findings suggest that GHCU plays a crucial role in the regulation of leaf flattening through facilitating cell-to-cell trafficking of KNGH1 and hence influencing the auxin response level.

Comparative transcriptomic analysis of rubber (Hevea brasiliensis) clones differing in disease tolerance provides novel insights into the disease response associated with abnormal leaf fall disease caused by Phytophthora meadii

Rubber is an important industrial commodity and Hevea brasiliensis is the commercial source of natural rubber (NR). Abnormal Leaf Fall (ALF) disease caused by Phytophthora meadii is one of the major diseases affecting yield of NR. Understanding Hevea-Phytophthora interaction mechanism is essential to manage this devastating disease. Transcriptome analysis was employed to gain an understanding on the molecular mechanisms that confer resistance to Phytophthora that would facilitate development of disease-tolerant Hevea clones. As our current understanding of Hevea-Phytophthora interaction is limited, an attempt was made to analyze and compare the transcriptomes of tolerant and susceptible clones of H. brasiliensis in response to P. meadii infection (in healthy and Phytophthora challenge-inoculated condition) during early, middle and late phases of infection by using the RNA-Seq technology. The transcriptome sequencing generated an average of 19,000 differentially expressed genes (DEGs) in both clones. When comparing the tolerant clone to the susceptible clone, there was an overall increase in the number of DEGs and a progression in the number of DEGs, indicating a more active engagement of transcriptional activity in the tolerant clone. Functional annotation, KEGG analysis and enrichment analysis revealed that the major pathways enriched for the DEGs were involved in plant hormone signal transduction and plant-pathogen interaction. An analysis of DEGs during each phase showed that the most active transcripts involved in defense mechanisms were activated during mid-phase and in the case of susceptible clone least transcripts were induced during mid-phase than in any other phase in both the clones. Many DEGs belonging to the MYB, NAC, WRKY and AP2/ERF transcription factor families were observed in both clones. Comparative studies revealed their differential involvement in tolerant and susceptible clones against Phytophthora infection. These findings may facilitate further exploration of Hevea-Phytophthora interaction and in cloning for defense related genes.

Comprehensive phytohormone metabolomic and transcriptomic analysis of tobacco (Nicotiana tabacum) infected by tomato spotted wilt virus (TSWV)

Tomato spotted wilt virus (TSWV) is ranked among the top 10 most destructive viruses globally. It results in abnormal leaf growth, stunting, and even death, significantly affecting crop yield and quality. Phytohormones play a crucial role in regulating plant-virus interactions. However, there is still limited research on the effect of TSWV on phytohormone levels, particularly growth hormones and genes involved in the phytohormone pathway. In our study, we combined phytohormone metabolomics and transcriptomics to examine the impact of TSWV infection on phytohormone content and gene expression profile. Metabolomic results showed that 41 metabolites, including major phytohormones and their precursors and derivatives were significantly altered after 14 days of TSWV inoculation tobacco plants cvK326, with 31 being significantly increased and 10 significantly reduced. Specifically, the levels of abscisic acid (ABA) and jasmonoyl-isoleucine (JA-Ile) were significantly reduced. The levels of indole-3-acetic acid (IAA) have remained unchanged. However, the levels of cytokinin isopentenyladenine (iP) and salicylic acid (SA) significantly increased. The transcriptome analysis revealed 2,746 genes with significant changes in expression. Out of these, 1,072 genes were significantly downregulated, while 1,674 genes were significantly upregulated. Among them, genes involved in ABA synthesis and signaling pathways, such as 9-cis-epoxycarotenoid dioxygenase (NCED), protein phosphatase 2C (PP2C), serine/threonine-protein kinase (SnRK2), and abscisic acid responsive element binding factor (ABF), exhibited significant downregulation. Additionally, expression of the lipoxygenase gene LOX, Jasmonate ZIM domain-containing protein gene JAZ, and transcription factor gene MYC were significantly down-regulated. In the cytokinin pathway, while there were no significant changes in the expression of the cytokinin synthesis genes, a significant downregulation of transcriptionally active factor type-B response regulators (type-B RRs) was observed. In terms of SA synthesis and signaling pathways, the isochorismate synthase gene ICS1 and the pathogenesis-related gene PR1 were significantly upregulated. These results can strengthen the theoretical foundation for understanding the interaction between TSWV and tobacco and provide new insights for the future prevention and control of TSWV.

Anatomical and physiological consequences of beech leaf disease in Fagus grandifolia L.

AbstractBeech leaf disease (BLD) is a foliar disease of American Beech (Fagus grandifolia L.) causally linked to the nematode Litylenchus crenatae mccannii and has rapidly spread throughout central and northeastern North America. This study aimed to characterize the anatomical and physiological differences between asymptomatic and symptomatic leaves to provide evidence for a mechanistic link between abnormal leaf development associated with BLD and the long‐term decline of affected trees. We found that leaf mass per area (LMA) and leaf thickness were 45% and 249% higher in symptomatic regions, respectively. The difference in leaf thickness was largely attributable to the spongy mesophyll being 410% thicker in symptomatic as compared with asymptomatic regions of the leaves, but palisade mesophyll and abaxial epidermal tissues were also thicker in symptomatic regions. While major vein density was not significantly different, minor vein density was significantly lower in symptomatic regions, suggesting that the effects on leaf development occur after the formation and initial development of first‐ and second‐order veins. Stomatal density was also lower in symptomatic leaves. Maximum photosynthetic rates were approximately 61% lower in symptomatic leaves and respiration rate increased as the percentage of affected leaf tissue increased. Collectively, our data show reduced photosynthetic capacity, increased respiration rates, and higher leaf construction costs, which will likely have a negative, long‐term impact on the carbon balance of trees affected by BLD.

Ectopic expression of PmGRF7 isolated from Japanese apricot in tomato leads to seed sterility

The role of GRF (Growth-regulating Factor) in regulating plant growth and development has been studied, but its potential role in regulating seed fertility remains unclear. PmGRF7 overexpression in tomatoes altered leaf morphology and pollen activity. Most importantly, genetically modified tomato plants had no offspring. PmGRF7 overexpression resulted in abnormal leaf morphology, and transgenic plants that produced seedless fruit or seeds only showed signs of abortion. Pollination of genetically modified tomatoes with wild-type tomato pollen still produced sterile seeds. Further research using bioinformatics tools and physiological index measurement showed that PmGRF7 induced a hormone pathway leading to seed sterility, and at the same time, it caused differential genes to be induced in the plant hormone signal transduction pathway, ribosome pathway, sphingolipid metabolism and other related pathways, leading to morphogenesis in plant leaves. Collectively, our research findings reveal a potential new mechanism by which PmGRF7 regulates seed fertility and leaf growth, and these findings are of potential application in molecular plant breeding.

Curling Leaf 1, Encoding a MYB-Domain Protein, Regulates Leaf Morphology and Affects Plant Yield in Rice.

The leaf is the main site of photosynthesis and is an important component in shaping the ideal rice plant architecture. Research on leaf morphology and development will lay the foundation for high-yield rice breeding. In this study, we isolated and identified a novel curling leaf mutant, designated curling leaf 1 (cl1). The cl1 mutant exhibited an inward curling phenotype because of the defective development of sclerenchymatous cells on the abaxial side. Meanwhile, the cl1 mutant showed significant reductions in grain yield and thousand-grain weight due to abnormal leaf development. Through map-based cloning, we identified the CL1 gene, which encodes a MYB transcription factor that is highly expressed in leaves. Subcellular localization studies confirmed its typical nuclear localization. Transcriptome analysis revealed a significant differential expression of the genes involved in photosynthesis, leaf morphology, yield formation, and hormone metabolism in the cl1 mutant. Yeast two-hybrid assays demonstrated that CL1 interacts with alpha-tubulin protein SRS5 and AP2/ERF protein MFS. These findings provide theoretical foundations for further elucidating the mechanisms of CL1 in regulating leaf morphology and offer genetic resources for practical applications in high-yield rice breeding.

First Report of Nothophoma quercina Causing Brown Spot Disease on 'Huanghua' Pear (Pyrus pyrifolia Nakai) in China.

Pear (Pyrus L.) is an important fruit tree in China, which has the largest cultivation area and yield in the world (Jia et al. 2021). In June 2022, brown spot symptoms were observed on 'Huanghua' pear (Pyrus pyrifolia Nakai, cv. Huanghua) leaves in the germplasm garden of Anhui Agricultural University (High Tech Agricultural Garden), Anhui, Hefei, China. The disease incidence was approximately 40% according to the percentage of diseased leaves among 300 leaves (50 leaves each were obtained from 6 plants). Initially, small, brown, round to oval lesions appeared on the leaves, the spots were gray in the central, and surrounded by brown to black margins. These spots rapidly enlarged, eventually causing abnormal leaf defoliation. To isolate the brown spot pathogen, symptomatic leaves were harvested, washed with sterile water, surface-sterilized with 75% ethanol for 20 s, and washed 3-4 times with sterile water. Leaf fragments were placed onto PDA medium and incubated at 25°C for 7 days to obtain isolates. The colonies exhibited white to pale gray aerial mycelium and reached a diameter of 62 mm after 7 days of incubation. Conidiogenous cells were characterized as phialides, and exhibited a doliform to ampulliform shape. Conidia displayed various shapes and sizes, ranging from subglobose to oval or obtuse, with thin walls, aseptate hyphae, and a smooth surface. They measured 4.2-7.9 × 3.1-5.5 μm in diameter. These morphologies were similar to Nothophoma quercina as reported previously (Bai et al. 2016; Kazerooni et al. 2021). For molecular analysis, the internal transcribed spacers (ITS), beta-tubulin (TUB2), and actin (ACT) regions were amplified using the primers ITS1/ITS4, Bt2a/Bt2b, and ACT-512F/ACT-783R respectively. The sequences of ITS, TUB2, and ACT were deposited in GenBank (accession numbers: OP554217, OP595395, and OP595396, respectively). A nucleotide blast search revealed high homology with N. quercina sequences: MH635156 (ITS: 541/541, 100%), MW672036.1 (TUB2: 343/346, 99%), FJ426914.1 (ACT: 242/262, 92%). A phylogenetic tree was constructed with ITS, TUB2 and ACT sequences based on neighbor-joining method using MEGA-X software, which showed the highest similarity with N. quercina. To confirm the pathogenicity, the leaves of three healthy plants were sprayed with spore suspension (106 conidia/mL), whereas control leaves were prayed with sterile water. The inoculated plants were covered with plastic bags and cultured in a growth chamber (90% relative humidity) at 25°C. Typical disease symptoms appeared on the inoculated leaves after 7-10 days, whereas no symptoms were observed on the control leaves. The same pathogen was re-isolated from the diseased leaves, according with Koch's postulates. Therefore, based on morphological and phylogenetic tree analyses, we confirmed that the causal organism for brown spot disease was N. quercina fungus (Chen et al. 2015; Jiao et al. 2017). To our knowledge, this is the first report of brown spot disease caused by N. quercina on 'Huanghua' pear leaves in China.

A large deletion conferring pale green leaves of maize

BackgroundThe structural basis of chloroplast and the regulation of chloroplast biogenesis remain largely unknown in maize. Gene mutations in these pathways have been linked to the abnormal leaf color phenotype observed in some mutants. Large scale structure variants (SVs) are crucial for genome evolution, but few validated SVs have been reported in maize and little is known about their functions though they are abundant in maize genomes.ResultsIn this research, a spontaneous maize mutant, pale green leaf-shandong (pgl-sd), was studied. Genetic analysis showed that the phenotype of pale green leaf was controlled by a recessive Mendel factor mapped to a 156.8-kb interval on the chromosome 1 delineated by molecular markers gy546 and gy548. There were 7 annotated genes in this interval. Reverse transcription quantitative PCR analysis, SV prediction, and de novo assembly of pgl-sd genome revealed that a 137.8-kb deletion, which was verified by Sanger sequencing, might cause the pgl-sd phenotype. This deletion contained 5 annotated genes, three of which, including Zm00001eb031870, Zm00001eb031890 and Zm00001eb031900, were possibly related to the chloroplast development. Zm00001eb031870, encoding a Degradation of Periplasmic Proteins (Deg) homolog, and Zm00001eb031900, putatively encoding a plastid pyruvate dehydrogenase complex E1 component subunit beta (ptPDC-E1-β), might be the major causative genes for the pgl-sd mutant phenotype. Plastid Degs play roles in protecting the vital photosynthetic machinery and ptPDCs provide acetyl-CoA and NADH for fatty acid biosynthesis in plastids, which were different from functions of other isolated maize leaf color associated genes. The other two genes in the deletion were possibly associated with DNA repair and disease resistance, respectively. The pgl-sd mutation decreased contents of chlorophyll a, chlorophyll b, carotenoids by 37.2%, 22.1%, and 59.8%, respectively, and led to abnormal chloroplast. RNA-seq revealed that the transcription of several other genes involved in the structure and function of chloroplast was affected in the mutant.ConclusionsIt was identified that a 137.8-kb deletion causes the pgl-sd phenotype. Three genes in this deletion were possibly related to the chloroplast development, which may play roles different from that of other isolated maize leaf color associated genes.

The nuclear-localized RNA helicase 13 is essential for chloroplast development in Arabidopsis thaliana.

The chloroplast is a semi-autonomous organelle with a double membrane structure, and its structural stability is a prerequisite for its correct function. Chloroplast development is regulated by known nuclear-encoded chloroplast proteins or proteins encoded within the chloroplast itself. However, the mechanism of chloroplast development regulated by other organelles remains largely unknown. Here, we report that the nuclear-localized DEAD-box RNA helicase 13 (RH13) is essential for chloroplast development in Arabidopsis thaliana. RH13 is widely expressed in tissues and localized to the nucleolus. A homozygous rh13 mutant shows abnormal chloroplast structure and leaf morphogenesis. Proteomic analysis showed that the expression levels of photosynthesis-related proteins in chloroplasts were reduced due to loss of RH13. Furthermore, RNA-sequencing and proteomics data revealed decreases in the expression levels of these chloroplast-related genes, which undergo alternative splicing events in the rh13 mutant. Taken together, we propose that nucleolus-localized RH13 is critical for chloroplast development in Arabidopsis.

Quantitative estimation of organ-scale phenotypic parameters of field crops through 3D modeling using extremely low altitude UAV images

Plant phenotypic parameters provide key information in modern crop breeding. However, the rapid and accurate estimation of organ-scale phenotypic parameters remains a challenge. In this context, the present study proposed a novel methodology for the automatic quantification of the organ-scale parameters of field crops using the unmanned aerial vehicle (UAV) platform. First, a lightweight UAV was employed to capture the multi-view and high-resolution image sequences of field crops at an extremely-low flight altitude. Subsequently, based on these image sequences, point cloud reconstruction of the canopy was conducted. Next, the geometrical model of individual leaves was reconstructed using the modified Crust algorithm and optimized by abnormal facet elimination and leaf surface repair. Finally, individual leaf phenotypic parameters were calculated based on the reconstructed geometrical models. The method was evaluated by comparing the calculated parameters with actual measurements. The calculated values for leaf length, maximum leaf width, and leaf area were in good agreements with the measured values (maize: R2 > 0.97 for all parameters, RMSE for length, width, and leaf area was 2.6 cm, 0.4 cm, and 33.2 cm2, respectively; soybean: R2 > 0.85 for all parameters, RMSE for the counterparts was 0.3 cm, 0.5 cm, and 2.3 cm2, respectively; tobacco: R2 > 0.89 for all parameters, RMSE for the counterparts was 4.1 cm, 1.4 cm, and 42.6 cm2, respectively). The methodology based on extremely-low altitude UAV images has promising prospects in the crop breeding program for the automatic acquisition of fine organ-scale parameters with high efficiency.

A Chronological Study on Grapevine Leafroll-Associated Virus 2 in Australia.

Grapevine leafroll disease affects the health status of grapevines worldwide. Most studies in Australia have focused on grapevine leafroll-associated viruses 1 and 3, while little attention has been given to other leafroll virus types, in particular, grapevine leafroll-associated virus 2 (GLRaV-2). A chronological record of the temporal occurrence of GLRaV-2 in Australia since 2001 is reported. From a total of 11,257 samples, 313 tested positive, with an overall incidence of 2.7%. This virus has been detected in 18 grapevine varieties and Vitis rootstocks in different regions of Australia. Most varieties were symptomless on their own roots, while Chardonnay showed a decline in virus-sensitive rootstocks. An isolate of GLRaV-2, on own-rooted Vitis vinifera cv. Grenache, clone SA137, was associated with severe leafroll symptoms after veraison with abnormal leaf necrosis. The metagenomic sequencing results of the virus in two plants of this variety confirmed the presence of GLRaV-2, as well as two inert viruses, grapevine rupestris stem pitting-associated virus (GRSPaV) and grapevine rupestris vein feathering virus (GRVFV). No other leafroll-associated viruses were detected. Among the viroids, hop stunt viroid and grapevine yellow speckle viroid 1 were detected. Of the six phylogenetic groups identified in GLRaV-2, we report the presence of four groups in Australia. Three of these groups were detected in two plants of cv. Grenache, without finding any recombination event. The hypersensitive reaction of certain American hybrid rootstocks to GLRaV-2 is discussed. Due to the association of GLRaV-2 with graft incompatibility and vine decline, the risk from this virus in regions where hybrid Vitis rootstocks are used cannot be overlooked.

Research on cassava disease classification using the multi-scale fusion model based on EfficientNet and attention mechanism.

Cassava disease is one of the leading causes to the serious decline of cassava yield. Because it is difficult to identify the characteristics of cassava disease, if not professional cassava growers, it will be prone to misjudgment. In order to strengthen the judgment of cassava diseases, the identification characteristics of cassava diseases such as different color of cassava leaf disease spots, abnormal leaf shape and disease spot area were studied. In this paper, deep convolutional neural network was used to classify cassava leaf diseases, and image classification technology was used to recognize and classify cassava leaf diseases. A lightweight module Multi-scale fusion model (MSFM) based on attention mechanism was proposed to extract disease features of cassava leaves to enhance the classification of disease features. The resulting feature map contained key disease identification information. The study used 22,000 cassava disease leaf images as a data set, including four different cassava leaf disease categories and healthy cassava leaves. The experimental results show that the cassava leaf disease classification model based on multi-scale fusion Convolutional Neural Network (CNN) improves EfficientNet compared with the original model, with the average recognition rate increased by nearly 4% and the average recognition rate up to 88.1%. It provides theoretical support and practical tools for the recognition and early diagnosis of plant disease leaves.

Insight into the formation of trumpet and needle-type leaf in Ginkgo biloba L. mutant.

The leaf type of a plant determines its photosynthetic efficiency and adaptation to the environment. The normal leaves of modern Ginkgo biloba, which is known as a "living fossil" in gymnosperm, evolved from needle-like to fan-shaped with obvious dichotomous venation. However, a newly discovered Ginkgo variety "SongZhen" have different leaf types on a tree, including needle-, trumpet-, strip-, and deeply split fan-shaped leaves. In order to explore the mechanism in forming these leaf types, the microscopy of different leaf types and transcriptome analysis of apical buds of branches with normal or abnormal leaves were performed. We found that the normal leaf was in an intact and unfolded fan shape, and the abnormal leaf was basically split into two parts from the petiole, and each exhibited different extent of variation. The needle-type leaves were the extreme, having no obvious palisade and spongy tissues, and the phloem cells were scattered and surrounded by xylem cells, while the trumpet-type leaves with normal vascular bundles curled inward to form a loop from the abaxial to adaxial side. The other type of leaves had the characteristics among needle-type, trumpet-type, or normal leaves. The transcriptome analysis and quantitative PCR showed that the genes related to abaxial domain were highly expressed, while the adaxial domain promoting genes were decreasingly expressed in abnormal-type leaf (ANL) buds and abnormal leaves, which might lead to the obvious abaxialized leaves of "SongZhen." In addition, the low expression of genes related to leaf boundary development in ANL buds indicated that single- or double-needle (trumpet) leaves might also be due to the leaf tissue fusion. This study provides an insight into the mechanism of the development of the abnormal leaves in "SongZhen" and lays a foundation for investigating the molecular mechanism of the leaf development in gymnosperms.

Association of phytoplasmas with a new disease of potato crops in cundinamarca, Colombia

Cundinamarca is the major potato producer region in Colombia. Potato growers first reported the presence of new symptoms affecting commercial potato fields in Cundinamarca as far back as 2012. These included foliage yellowing, curling on the upper leaf side and purple margins, occasionally with purple coloration of the underside veins, excessive shoot proliferation, abnormally short or long internodes and abnormal leaf development. Plants had a distorted pattern of growth suggesting a possible phytoplasma cause. Phytoplasmas were detected by nested PCR of the phytoplasma 16S rRNA gene and we confirmed the presence of ‘Candidatus Phytoplasma asteris’ (group 16SrI) and ‘Candidatus Phytoplasma fraxini’ (16SrVII) by RFLP and sequencing of samples from commercial plots. Of 152 symptomatic and non-symptomatic potato plants tested, 33.5% were positive for one of the identified phytoplasmas or both (mixed infections). Phytoplasmas were also observed by electron microscopy in symptomatic plants. Both phytoplasmas were transmitted in tubers produced by infected plants and by the insect vector Exitianus atratus (Hemiptera: Cicadellidae) to healthy test potato plants. Daughter plants from infected tubers and the healthy plants inoculated via the insect vector developed the same symptoms observed in field grown potato plants, in single and mixed infections of both phytoplasmas. Our results confirm the presence of a new disease of potato in Cundinamarca, Colombia, which we named yellowing and rolling-leaf phytoplasma (fitoplasma del amarillamiento y enrrollamiento de hojas).

DRB2 Modulates Leaf Rolling by Regulating Accumulation of MicroRNAs Related to Leaf Development in Rice.

As an important agronomic trait in rice (Oryza sativa), moderate leaf rolling helps to maintain the erectness of leaves and minimize shadowing between leaves, leading to improved photosynthetic efficiency and grain yield. However, the molecular mechanisms underlying rice leaf rolling still need to be elucidated. Here, we isolated a rice mutant, rl89, showing adaxially rolled leaf phenotype due to decreased number and size of bulliform cells. We confirmed that the rl89 phenotypes were caused by a single nucleotide substitution in OsDRB2 (LOC_Os10g33970) gene encoding DOUBLE-STRANDED RNA-BINDING2. This gene was constitutively expressed, and its encoded protein was localized to both nucleus and cytoplasm. Yeast two-hybrid assay showed that OsDRB2 could interact with DICER-LIKE1 (DCL1) and OsDRB1-2 respectively. qRT-PCR analysis of 29 related genes suggested that defects of the OsDRB2-miR166-OsHBs pathway could play an important role in formation of the rolled leaf phenotype of rl89, in which OsDRB2 mutation reduced miR166 accumulation, resulting in elevated expressions of the class III homeodomain-leucine zipper genes (such as OsHB1, 3 and 5) involved in leaf polarity and/or morphology development. Moreover, OsDRB2 mutation also reduced accumulation of miR160, miR319, miR390, and miR396, which could cause the abnormal leaf development in rl89 by regulating expressions of their target genes related to leaf development.

Disruption of tomato TGS machinery by ToLCNDV causes reprogramming of vascular tissue-specific TORNADO1 gene expression.

Vascular development-related TRN1 transcription is suppressed by cytosine methylation in fully developed leaves of tomato. ToLCNDV infection disrupts methylation machinery and reactivates TRN1 expression - likely causing abnormal leaf growth pattern. Leaf curl disease of tomato caused by tomato leaf curl New Delhi virus (ToLCNDV) inflicts huge economical loss. Disease symptoms resemble leaf developmental defects including abnormal vein architecture. Leaf vein patterning-relatedTORNADO1gene's (SlTRN1) transcript level is augmented in virus-infected leaves. To elucidate the molecular mechanism of the upregulation ofSlTRN1in vivo, we have deployedSlTRN1promoter-reporter transgenic tomato plants and investigated the gene's dynamic expression pattern in leaf growth stages and infection. Expression of the gene was delimited in the vascular tissues and suppressed in fully developed leaves. WRKY16 transcription factor readily activated SlTRN1 promoter in varied sized leaves and upon virus infection, while silencing of WRKY16 gene resulted in dampened promoter activity. Methylation-sensitive PCR analyses confirmed the accumulation of CHH methylation at multiple locations in theSlTRN1promoter in older leaves. However, ToLCNDV infection reverses the methylation status and restores expression level in the leaf vascular bundle. The virus dampens the level of key maintenance and de novo DNA methyltransferases SlDRM5, SlMET1, SlCMT2 with concomitant augmentation of two DNA demethylases, SlDML1 and SlDML2 levels inSlTRN1promoter-reporter transgenics. Transient overexpression of SlDML2 mimics the virus-induced hypomethylation state of theSlTRN1promoter in mature leaves, while silencing of SlDML2 lessens promoter activity. Furthermore, in line with the previous studies, we confirm the crucial role of viral suppressors of RNA silencing AC2 and AC4 proteins in promoting DNA demethylation and directing it to restore activated transcription ofSlTRN1. Unusually elevated expression of SlTRN1 may negatively impact normal growth of leaves.