Plant Direct最新文献

RNA interference (RNAi) is a crucial mechanism in immunity against infectious microbes through the action of DICER-LIKE (DCL) and ARGONAUTE (AGO) proteins. In the case of the taxonomically diverse fungal pathogen Botrytis cinerea and the oomycete Hyaloperonospora arabidopsidis, plant DCL and AGO proteins have proven roles as negative regulators of immunity, suggesting functional specialization of these proteins. To address this aspect in a broader taxonomic context, we characterized the colonization pattern of an informative set of DCL and AGO loss-of-function mutants in Arabidopsis thaliana upon infection with a panel of pathogenic microbes with different lifestyles, and a fungal mutualist. Our results revealed that, depending on the interacting pathogen, AGO1 acts as a positive or negative regulator of immunity, while AGO4 functions as a positive regulator. Additionally, AGO2 and AGO10 positively modulated the colonization by a fungal mutualist. Therefore, analyzing the role of RNAi across a broader range of plant-microbe interactions has identified previously unknown functions for AGO proteins. For some pathogen interactions, however, all tested mutants exhibited wild-type-like infection phenotypes, suggesting that the roles of AGO and DCL proteins in these interactions may be more complex to elucidate.

Thylakoid membranes in chloroplasts and cyanobacteria harbor the multisubunit protein complexes that catalyze the light reactions of photosynthesis. In plant chloroplasts, the thylakoid membrane system comprises a highly organized network with several subcompartments that differ in composition and morphology: grana stacks, unstacked stromal lamellae, and grana margins at the interface between stacked and unstacked regions. The localization of components of the photosynthetic apparatus among these subcompartments has been well characterized. However, less is known about the localization of proteins involved in the biogenesis and repair of the photosynthetic apparatus, the partitioning of proteins between two recently resolved components of the traditional margin fraction (refined margins and curvature), and the effects of light on these features. In this study, we analyzed the partitioning of numerous thylakoid biogenesis and repair factors among grana, curvature, refined margin, and stromal lamellae fractions of Arabidopsis thylakoid membranes, comparing the results from illuminated and dark-adapted plants. Several proteins previously shown to localize to a margin fraction partitioned in varying ways among the resolved curvature and refined margin fractions. For example, the ALB3 insertase and FtsH protease involved in photosystem II (PSII) repair were concentrated in the refined margin fraction, whereas TAT translocon subunits and proteins involved in early steps in photosystem assembly were concentrated in the curvature fraction. By contrast, two photosystem assembly factors that facilitate late assembly steps were depleted from the curvature fraction. The enrichment of the PSII subunit OE23/PsbP in the curvature fraction set it apart from other PSII subunits, supporting the previous conjecture that OE23/PsbP assists in PSII biogenesis and/or repair. The PSII assembly factor PAM68 partitioned differently among thylakoid fractions from dark-adapted plants and illuminated plants and was the only analyzed protein to convincingly do so. These results demonstrate an unanticipated spatial heterogeneity of photosystem biogenesis and repair functions in thylakoid membranes and reveal the curvature fraction to be a focal point of early photosystem biogenesis.

Screening a transposon-mutagenized soybean population led to the discovery of a recessively inherited chlorotic phenotype. This "y24" phenotype results in smaller stature, weaker stems, and a smaller root system. Genome sequencing identified 15 candidate genes with mutations likely to result in a loss of function. Amplicon sequencing of a segregating population was then used to narrow the list to a single candidate mutation, a single-base change in Glyma.07G102300 that disrupts splicing of the second intron. Single cell transcriptomic profiling indicates that this gene is expressed primarily in mesophyll cells, and RNA sequencing data indicate that it is upregulated in germinating seedlings by cold stress. Previous studies have shown that mutations to Os05g34040, the rice ortholog of Glyma.07G102300, produced a chlorotic phenotype that was more pronounced in cool temperatures. Growing soybean y24 mutants at lower temperatures also resulted in a more severe phenotype. In addition, transgenic expression of wild-type Glyma.07G102300 in the knockout mutant of the Arabidopsis ortholog At4930720 rescues the chlorotic phenotype, further supporting the hypothesis that the mutation in Glyma.07G102300 is causal of the y24 phenotype. The variant analysis strategy used to identify the genes underlying this phenotype provides a template for the study of other soybean mutants.

Interdisciplinarity is used to integrate and synthesize new research directions between scientific domains, but it is not the only means by which to generate novelty by bringing diverse perspectives together. Internationality draws upon cultural and linguistic diversity that can potentially impact interdisciplinarity as well. We created an interdisciplinary class originally intended to bridge computational and plant science that eventually became international in scope, including students from the United States and Mexico. We administered a survey over 4 years designed to evaluate student expertise. The first year of the survey included only US students and demonstrated that biology and computational student groups have distinct expertise but can learn the skills of the other group over the course of a semester. Modeling of survey responses shows that biological and computational science expertise is equally distributed between US and Mexico student groups, but that nonetheless, these groups can be predicted based on survey responses due to subspecialization within each domain. Unlike interdisciplinarity, differences arising from internationality are mostly static and do not change with educational intervention and include unique skills such as working across languages. We end by discussing a distinct form of interdisciplinarity that arises through internationality and the implications of globalizing research and education efforts.

Burley tobacco, a chlorophyll-deficient mutant with impaired nitrogen use efficiency (NUE), generally requires three to five times more nitrogen fertilization than flue-cured tobacco to achieve a comparable yield, which generates serious environmental pollution and negatively affects human health. Therefore, exploring the mechanisms underlying NUE is an effective measure to reduce environmental pollution and an essential direction for burley tobacco plant improvement. Physiological and genetic factors affecting tobacco NUE were identified using two tobacco genotypes with contrasting NUE in hydroponic experiments. Nitrogen use inefficient genotype (TN90) had lower nitrogen uptake and transport efficiencies, reduced leaf and root biomass, lower nitrogen assimilation and photosynthesis capacity, and lower nitrogen remobilization ability than the nitrogen use efficient genotype (K326). Transcriptomic analysis revealed that genes associated with photosynthesis, carbon fixation, and nitrogen metabolism are implicated in NUE. Three nitrate transporter genes in the leaves (NPF2.11, NPF2.13, and NPF3.1) and three nitrate transporter genes (NPF6.3, NRT2.1, and NRT2.4) in roots were down-regulated by nitrogen starvation, all of which showed lower expression in TN90 than in K326. In addition, the protein-protein interaction (PPI) network diagram identified eight key genes (TPIP1, GAPB, HEMB, PGK3, PSBO, PSBP2, PSAG, and GLN2) that may affect NUE. Less advantageous changes in nitrogen uptake, nitrogen assimilation in combination with nitrogen remobilization, and maintenance of photosynthesis in response to nitrogen deficiency led to a lower NUE in TN90. The key genes (TPIP1, GAPB, PGK3, PSBO, PSBP2, PSAG, and GLN2) were associated with improving photosynthesis and nitrogen metabolism in tobacco plants grown under N-deficient conditions.

Anthropogenic increase in carbon dioxide (CO₂) affects plant physiology. Plant responses to elevated CO₂ typically include: (1) enhanced photosynthesis and increased primary productivity due to carbon fertilization and (2) suppression of leaf transpiration due to CO₂-driven decrease in stomatal conductance. The combined effect of these responses on the total plant transpiration and on evapotranspiration (ET) has a wide range of implications on local, regional, and global hydrological cycles, and thus needs to be better understood. Here, we investigated the net effect of CO₂-driven perennial ryegrass (Lolium perenne) physiological responses on transpiration and evapotranspiration by integrating physiological and hydrological (water budget) methods, under a controlled environment. Measurements of the net photosynthetic rate, stomatal conductance, transpiration rate, leaf mass per area, aboveground biomass, and water balance components were recorded. Measured variables under elevated CO₂ were compared with those of ambient CO₂. As expected, our results show that elevated CO₂ significantly decreases whole-plant transpiration rates (38% lower in the final week) which is a result of lower stomatal conductance (57% lower in the final week) despite a slight increase in aboveground biomass. Additionally, there was an overall decline in evapotranspiration (ET) under elevated CO₂, indicating the impact of CO₂-mediated suppression of transpiration on the overall water balance. Although studies with larger sample sizes are needed for more robust conclusions, our findings have significant implications for global environmental change. Reductions in ET from ryegrass-dominated grasslands and pastures could increase soil moisture and groundwater recharge, potentially leading to increased surface runoff and flooding.

Sequence-specific endonucleases have been key to the study of the mechanisms and control of DNA double-strand break (DSB) repair and recombination, and the availability of CRISPR-Cas nucleases over the last decade has driven rapid progress in the understanding and application of targeted recombination in many organisms, including plants. We present here an analysis of recombination at targeted chromosomal 5' overhang DSB generated by the FnCas12a endonuclease in the plant, Arabidopsis thaliana. The much-studied Cas9 nuclease cleaves DNA to generate blunt-ended DSBs, but relatively less is known about the repair of other types of breaks, such as those with 5'-overhanging ends. Sequencing the repaired breaks clearly shows that the majority of repaired DSB carry small deletions and are thus repaired locally by end-joining recombination, confirmed by Nanopore sequencing of larger amplicons. Paired DSBs generate deletions at one or both cut-sites, as well as deletions and reinsertions of the deleted segment between the two cuts, visible as inversions. While differences are seen in the details, overall the deletion patterns are similar between repair at single-cut and double-cut events, notwithstanding the fact that only the former involve cohesive DNA overhangs. A strikingly different repair pattern is however observed at breaks flanked by direct repeats. This change in sequence context results in the presence of a major alternative class of repair events, corresponding to highly efficient repair by single-strand annealing recombination.

The flowering time and plant architecture of Brassica napus were significantly associated with yield. In this study, we found that the BREVIPEDICELLUS/KNAT1(BP) gene regulated the flowering time and plant architecture of B. napus. However, the precise regulatory mechanism remains unclear. We cloned two homologous BP genes, BnaBPA03 and BnaBPC03, from B. napus Xiaoyun. The protein sequence analysis showed two proteins containing conserved domains KNOX I, KNOX II, ELK, and HOX of the KONX protein family. The CRISPR/Cas9 knockout lines exhibited early budding and flowering time, coupled with floral organ abscission earlier and a larger leaf angle. On the contrary, overexpression plants displayed a phenotype that was the inverse of these characteristics. Furthermore, we observed upregulation of gibberellin and ethylene biosynthesis genes, as well as floral integrator genes in knocked-out plants. The results revealed that BnaBPs play a role in flowering time, floral organ abscission, and leaf angle as well as germination processes mediated. Additionally, BnaBPs exerted an impact on the biosynthesis pathways of ethylene and GA.

C₃ photosynthesis can be complemented with a C₄ carbon concentrating mechanism (CCM) to minimize photorespiratory losses. C₄ photosynthesis is often more efficient than C₃ under steady-state conditions. However, the C₄ CCM depends on inter-cellular metabolite concentration gradients, which must increase following increases in light intensity and could decrease rates of C₄ photosynthesis under fluctuating light. Additionally, incomplete flux through photorespiration could prove beneficial to C₄ assimilation during light induction of the CCM. Here, we compare metabolic profiles in the closely related C₃ Flaveria robusta and C₄ Flaveria bidentis during a light transient from low to high light to determine if these non-steady state accumulation patterns provide insight to the induction of the metabolite gradients needed to drive C4 intermediate transport and if there is incomplete cycling of photorespiratory intermediates. In these C₃ and C₄ species, metabolite steady-state pool sizes suggest that C₄ transport acids maintain concentration gradients across the bundle sheath and mesophyll cell types under these light fluctuations. However, there was incomplete flux through photorespiration in the C₄ F. bidentis, which could reduce photorespiratory CO₂ loss via glycine decarboxylation and help maintain higher rates of assimilation during following induction periods.

Soybean (Glycine max L.) is an important leguminous plant, in which pests trigger significant damage every year. Important members of this community are insects with piercing-sucking mouthpart, especially the southern green stinkbug, Nezara viridula L.. This insect with its extraoral digestion causes visible alterations (morphological and color changes) in the seeds. We aimed to obtain precise information about the extent and nature of damage in soybeans caused by N. viridula using nondestructive imaging methods. Two infestation conditions were applied: one with controlled numbers of pests (six insects/15 pods) and another with naturally occurring pests (samples collected from the apical part of the plant and samples from whole plants). An intact control group was also included, resulting in four treatment groups. Seed samples were analyzed by computed tomography (CT) and image color analysis under laboratory conditions. According to our CT findings, the damage caused by N. viridula changed the radiodensity, volume, and shape (Solidity) of the soybean seeds during the pod-filling and maturing period. Radiodensity was significantly reduced in all three damaged categories compared to the intact sample; the mean radiodensity reduction range was 49-412 HU. The seed volume also decreased significantly (25%-80% decrease), with a threefold reduction for samples exposed to regulated damage compared to natural ones. The samples exposed to natural damage showed significant but minor reduction in solidity, while samples exposed to regulated damage showed a prominent decrease (~12%). Image color analysis showed that the damaged samples were well distinguishable, and the differences were statistically verifiable. The achieved data derived from our external and internal imaging approaches contribute to a better understanding of the internal chemical processes, and CT analysis helps to understand the alteration trends of the hidden structure of seeds caused by a pest. Our results can contribute to the development of a practically applicable system based on image analysis, which can identify lots damaged by insects.