Kaede for Detection of Protein Oligomerization

Photoconvertible fluorescent proteins such as Kaede are routinely used for tracking proteins, organelles, and whole cells. Kaede was the first identified photoconvertible fluorescent protein and has since become the most commonly used photoconvertible fluorescent protein in vertebrates. Kaede can be irreversibly converted from a green to a red fluo- rescent form upon UV/blue light irradiation and fluorescence of each form can be isolated separately by appropriate filter sets. Spectral properties of the Kaede forms allow F6rster resonance energy transfer （FRET） from the green form as donor to the red form as acceptor. As a sample containing oligomerized Kaede-containing proteins is exposed to UV or blue light, FRET first increases as green Kaede is converted to red and then decreases as the green donor becomes depleted. Thus, FRET information is potentially obtained from a number of independent measurements taken as photoconversion proceeds. We demonstrate here the application of this approach to detect homo-aggregation and conformational dynamics of plant pro- tein constructs. Structural alterations of 2-cys peroxiredoxin-Kaede were successfully detected depending on the redox state in living plant cells. Photoconversion was performed gradually and donor emission, acceptor emission, and FRET-derived sensitized acceptor emission were measured at each step of conversion. Since photoconvertible proteins have not been rou- tinely used in plants, two plasmids have been designed to facilitate plant applications. The plasmids allow either transient expression of Kaede-containing protein constructs in plant cells or Gateway cloning and stable transformation of plants.     

一种水稻卷叶性状的遗传分析

对水稻卷叶品种流岗卷叶粳与4个平展叶品种及1个卷叶标志基因(rl3)系的杂交或回交后代进行了考察。结果表明,流岗卷叶粳的卷叶特性以单基因不完全显性方式遗传; 该基因与rl3基因不等位,当rl3位点处于隐性纯合时两者以累加方式发生互作。这一等位基因可作标志基因使用,暂定名为Rl(t)。 Abstract:Liugangjuanyejing is a rolled leaf mutant of rice.A genetic study on the rolled-leaf character was carried out by crossing it with four flat-leaf cultivars and a genetic marker line (with a rolled-leaf allele rl3).The results showed that this character was controlled by an incomplete dominant gene which was non-allelic to rl3 locus and that there existed additive effect between the two loci when the rl3 locus was homozygous recessiveness.This new rolled-leaf allele was provisionally named as Rl(t) and could be used as a genetic marker of rice.     

Identification of a delayed leaf greening gene from a mutation of pummelo

Delayed greening of young leaves is an unusual phenomenon of plants in nature. Citrus are mostly evergreen tree species. Here, a natural mutant of “Guanxi” pummelo(Citrus maxima), which shows yellow leaves at the young stage, was characterized to identify the genes underlying the trait of delayed leaf greening in plants. A segregating population with this mutant as the seed parent and a normal genotype as the pollen parent was generated. Two DNA pools respectively from the leaves of segregating seedlings with extreme phenotypes of normal leaf greening and delayed leaf greening were collected for sequencing. Bulked segregant analysis(BSA) and In Del marker analysis demonstrated that the delayed leaf greening trait is governed by a 0.3 Mb candidate region on chromosome 6. Gene expression analysis further identified a key candidate gene(Citrus Delayed Greening gene 1, CDG1) in the 0.3 Mb region, which showed significantly differential expression between the genotypes with delayed and normal leaf greening phenotypes. There was a 67 bp In Del region difference in the CDG1 promoter and the In Del region contains a TATA-box element. Confocal laser-scanning microscopy revealed that the CDG1-GFP fusion protein signals were co-localized with the chloroplast signals in the protoplasts. Overexpression of CDG1 in tobacco and Arabidopsis led to the phenotype of delayed leaf greening. These results suggest that the CDG1 gene is involved in controlling the delayed leaf greening phenotype with important functions in chloroplast development.     

Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants: silencing of the tomato ferric chelate reductase gene, FRO1, as an example

Virus-induced gene silencing (VIGS) is potentially an attractive reverse-genetics tool for studies of plant gene function, but whether it is effective in silencing mineral nutritional-related genes in roots has not been demonstrated. Here we report on an efficient VIGS system that functions in tomato roots using a modified viral satellite DNA (DNAmβ) associated with Tomato yellow leaf curl China virus (TYLCCNV). A cDNA fragment of the ferric chelate reductase gene (FRO1) from tomato was inserted into the DNAmβ vector. Tomato roots agro-inoculated with DNAmβ carrying both a fragment of FRO1 and TYLCCNV used as a helper virus exhibited a significant reduction at the FRO1 mRNA level. As a consequence, ferric chelate reductase activity, as determined by visualization of the pink FeBPDS3 complex was significantly decreased. Our results clearly demonstrated that VIGS system can be employed to investigate gene function associated with plant nutrient uptake in roots.     

Bionic Leaf Simulating the Thermal Effect of Natural Leaf Transpiration

We proposed a kind of bionic leaf to simulate the thermal effect of leaf transpiration. The bionic leaf was firstly designed to be composed of a green coating, a water holding layer, a Composite Adsorbent （CS） layer and an adsorption-desorption rate controlling layer. A thermophysical model was established for the bionic leaf, and the dynamic simulation results reveal that the water holding layer is not necessary; a CS of high thermal conductivity should be selected as the CS layer; the adsorp- tion-desorption rate controlling layer could be removed due to the low adsorption-desorption rate of the CS; and when CaC12 mass fraction of the CS reaches 40%; the bionic leaf could simulate the dynamic thermal behavior of the natural leaf. Based on the simulation results, we prepared bionic leaves with different CaC12 content. The thermographies of the bionic leaf and the natural leaf were shot using the Infrared Thermal Imager. The measured average radiative temperature difference between the bionic and natural leaves is less than 1.0 ℃.     

Characterization and Molecular Mapping of a New Virescent Mutant in Rice

Plant leaves play a significant role in photosynthesis. Normal chloroplast development is critical for plant growth and yield performance. Defect of the chlorophyll in chloroplasts may cause abnormal leaf colors, such as yellow, white, or stripe. Chloroplasts have their own genomes encoding for about 100 genes that are essential for plastid protein synthesis and photosynthesis （Kanno and Hirai, 1993; Sato et al., 1999）. Moreover, over 3000 proteins encoded by plant nuclear genomes target to the chloroplasts and participate in the chloroplast development and/or photosynthesis. Hitherto, a number of plant genes, which encode for enzymes involved in chlorophyll biosynthetic pathways,     

Response of Solanum tuberosum to Myzus persicae infestation at different stages of foliage maturity

Young leaves of the potato Solanum tuberosum L. cultivar Kardal contain resistance factors to the green peach aphid Myzuspersicae （Sulzer） （Hemiptera： Aphididae） and normal probing behavior is impeded. However, M. persicae can survive and reproduce on mature and senescent leaves of the cv. Kardal plant without problems. We compared the settling ofM. persicae on young and old leaves and analyzed the impact of aphids settling on the plant in terms of gene expression. Settling, as measured by aphid numbers staying on young or old leaves, showed that after 21 h significantly fewer aphids were found on the young leaves. At earlier time points there were no difference between young and old leaves, suggesting that the young leaf resistance factors are not located at the surface level but deeper in the tissue. Gene expression was measured in plants at 96 h postinfestation, which is at a late stage in the interaction and in compatible interactions this is long enough for host plant acceptance to occur. In old leaves of cv. Kardal （compatible interaction）, M. persicae infestation elicited a higher number of differentially regulated genes than in young leaves. The plant response to aphid infestation included a larger number of genes induced than repressed, and the proportion of induced versus repressed genes was larger in young than in old leaves. Several genes changing expression seem to be involved in changing the metabolic state of the leaf from source to sink.     

Responses of nitrogen and phosphorus resorption from leaves and branches to long-term nitrogen deposition in a Chinese fir plantation北大核心CSCD

Aims Our objectives were to investigate differences in nutrient resorption between different plant organs (leaf and branch), among plants with different life spans (one-year old, two-year old and senesced), and under different duration of nitrogen (N) deposition treatments in a Chinese fir (Cunninghamia lanceolata) plantation. Methods The long-term N deposition experiment was conducted in a 12-year-old fir plantation of subtropical China. N deposition treatment was initiated in January 2004 until now, up-going 14 years. N deposition were designed at 4 levels of 0, 60, 120, and 240 kg·hm–2·a–1, indicated as N0, N1, N2, and N3, respectively, with 3 replicates for each treatment. The solution of CO(NH2)2 was sprayed on the forest floor each month. In the study, we measured N and phosphorus (P) concentrations and analyzed the pattern of nutrient resorption of mature and senescing leaves and branches. The different responses of needles N and P resorption after 7- and 14-year N deposition treatments were also compared. Important findings After 14 years of N deposition, (1) during the senescing process, leaf and branch C, N, and P content gradually decreased with increasing treatment duration, with higher content in leaf than in branch. N content decreased in the order of one-year old green leaf > two-year old green leaf > senescent leaf > one-year old living branch > two-year old living branch > senescent branch, and N3 > N2 > N1 > N0, with C:N showing the opposite trend. Senescent organs had higher C:N, N:P, and C:P than mature living organs. N deposition increased N, N:P, and C:P of mature living organs (except for the two-year old green leaf), while decreased P and C:N. (2) N resorption efficiency (REN) and P resorption efficiency (REP) of leaves and branches decreased gradually with increasing life span. REP was typically higher in leaf and branch than REN. Leaf had lower REN (28.12%) than branch (30.00%), but higher REP (45.82%) than branch (30.42%). A highly significant linear correlation existed between N:P and REN:REP in leaves and branches. (3) REN decreased but REP increased with the treatment duration of N deposition. The longer experimental duration (14 years) reduced REN by 9.85%, 3.17%, 11.71% under N1, N2, and N3 treatments, respectively, and increased REP by 71.98%, 42.25%, 9.60%, respectively, than the shorter treatment duration (7 years). In summary, the responses of essential nutrients resorption efficiency for different plant organs and life span varied with the levels and duration of N deposition treatment. REN:REP in leaf and branch were mostly driven by N:P of leaf and branch. The results highlight that nutrients resorption is significantly influenced by long-term N deposition. © Chinese Journal of Plant Ecology.     

Rock paper scissors: CRISPR/Cas9-mediated interference with geminiviruses in plants

<正>Geminiviruses are a group of circular single-stranded DNA viruses that constitute the largest family of plant viruses.Many diseases resulting from geminivirus infections, such as maize streak disease, cassava mosaic disease, tomato yellow leaf curl disease and cotton leaf curl diseases, cause significant problems in terms of economic losses, posing a serious threat to global crop productivity (Hanley-Bowdoin et al., 2013; Yang et al., 2019). As obligate intracellular     

煤矸石山上不同种植年限和生长期香根草各部位全氮含量及其分配比例的比较

为了探明香根草〔Vetiveria zizanioides(Linn.)Nash〕在尾矿生态系统恢复中的作用机制,以贵州省六盘水市大河煤矿煤矸石山种植3、6、8和13 a的香根草为研究对象,分别在返青期、快速生长期、成熟期和枯黄期对香根草根、茎、叶和全株的全氮含量及其分配比例进行了比较.结果表明:在种植3、6、8和13 a,香根草根、茎、叶和全株的全氮含量均在快速生长期最高,并显著高于其余生长期.随种植年限增加,返青期、成熟期和枯黄期根的全氮含量,枯黄期茎的全氮含量以及返青期全株的全氮含量均逐渐升高;返青期茎和叶的全氮含量逐渐降低;快速生长期根的全氮含量,快速生长期和成熟期茎的全氮含量以及快速生长期、成熟期和枯黄期叶和全株的全氮含量先升高后降低,并在种植8 a最高.在种植6、8和13 a,快速生长期根的全氮含量分配比例最低,而快速生长期叶的全氮含量分配比例却最高.随种植年限增加,返青期根的全氮含量分配比例逐渐升高,而返青期茎和叶的全氮含量分配比例却逐渐降低;在种植8 a,快速生长期、成熟期和枯黄期根的全氮含量分配比例以及枯黄期茎的全氮含量分配比例最低或较低,而快速生长期和成熟期茎的全氮含量分配比例以及快速生长期、成熟期和枯黄期叶的全氮含量分配比例最高.在相同种植年限和生长期,香根草茎的全氮含量及其分配比例明显低于根和叶;总体来看,返青期、成熟期和枯黄期根的全氮含量及其分配比例高于叶,而快速生长期根的全氮含量及其分配比例却低于叶.研究结果显示:随着种植年限增加和生长期推移,香根草能够合理分配氮素资源,使其在煤矸石山立足,据此认为,香根草可用于煤矸石山的生态恢复和植被重建,但种植年限不宜超过8a.     

Do aphids paint the tree red (or yellow)—can herbivore resistance or photoprotection explain colourful leaves in autumn?

We explored two mutually nonexclusive hypotheses on autumnal leaf colouration. The co-evolutionary hypothesis states that autumnal leaf colouration functions as a handicap signal to herbivorous insects, whereas the photoprotection hypothesis posits that plant pigments primarily protect the plant against cold-induced photoinhibition and enhance nutrient transfer. To contrast both hypotheses, we compared yellow and red leaf colouration in three groups of mountain ash (Sorbus aucuparia L.). Two montane groups of different age were characterised by low aphid numbers and low temperature, and a lowland group by high aphid numbers and high temperature. There were no consistent altitudinal differences in leaf colouration. Compared to young trees, adult trees developed fewer red but more yellow leaves at high altitude. In the lowland population, the development of red leaf colour was related to decreasing daytime temperature, whereas the appearance of yellow leaf colouration corresponded to the decreasing photoperiod. This is consistent with the photoprotection hypothesis. Individual differences in red and yellow leaf colouration were inversely correlated to the number of fruits, which might be interpreted as a trade-off between reproductive and protective commitment. Temperature effects explained variation in aphid numbers over time and leaf colouration explained aphid distribution on a given day. As predicted by the co-evolutionary hypothesis, strongly coloured individuals harboured fewer aphids than green or dull-coloured ones. Since decreasing temperature reduced the number of migrating aphids but induced red leaf colouration, these processes are not mutually fine-tuned, which likely restricts the potential for co-evolution between mountain ash and aphids.     

RAPD analysis of blast resistant somaclones from upland rice cultivar IAC 47 for genetic divergence

Seventeen somaclones of upland rice cultivar IAC 47 showing different plant types, and either resistance or susceptibility to leaf blast, were utilized for random amplified polymorphic DNA (RAPD) analysis. Somaclones exhibited differences in reaction to isolates of Pyricularia grisea. Two somaclones (SC02 and SC04) were resistant to all three field isolates of somaclones, while the cultivar IAC 47 was susceptible. The inheritance study of two distinct plant types, one with erect bright green leaves and the other with droopy yellow green leaves, showed that a single possibly different, dominant gene governs each plant type. Of 32 random decamer primers utilized, OPA02 and OPD02 detected polymorphisms between somaclones showing erect bright green leaves and droopy yellow green leaves. Reliable grouping exhibiting 80% similarity was achieved with 17 primers. Leaf blast resistance to race IC-2 of P. grisea was associated with the plant type of erect bright green leaves.     

Metabolism of [14C]trans-zeatin and [14C]benzyladenine by letached yellow, green and variegated leaves of Schefflera

[8-¹⁴C]Benzyladenine (BA) and [8-¹⁴C] trans-zeatin (tZ) were fed through the petiole to mature, detached green, yellow and variegated leaves of Schefflera arboricola. Recovery of radioactivity from the plant material ranged between 4.2 and 22.1%. More radioactivity was recovered when tZ was applied compared to BA. Green leaves or the green parts of variegated leaves yielded more radioactivity than the yellow leaf material. BA was metabolized much faster than the endogenous cytokinin tZ. It would appear that while lower amounts of radioactivity were present in yellow leaves, as well as in yellow parts of variegated leaves, the rate of cytokinin metabolism was nevertheless faster. Metabolites that were formed to a greater extent in these yellow parts were the nucleotides of both cytokinins. Currently it is not known whether or not cytokinins influence chlorophyll and other pigment development in chimeric variegated leaves.     

Transmission of the Monocarpic Senescence Signal via the Xylem in Soybean

During monocarpic senescence in soybean (Glycine max [L.] Merrill cv. Anoka) there is a remobilization of nitrogen from the leaves to the seeds, and it has been hypothesized that this loss of nitrogen from the leaves induces foliar yellowing. The phloem in a small segment of the petiole between the pods and the target leaf can be inactivated with a jet of steam. When a plant is depodded except for a single pod cluster in the center of the plant, the pod cluster induces yellowing of the nearest leaf even if the petiole contains a zone of dead phloem, whereas most of the rest of the plant remains green. The nitrogen content of these leaves with a dead phloem zone in their petioles does not decrease greatly, even though the leaves turn yellow. A similar treatment of a single leaf on a fully depodded plant (leaves stay green) does not cause that leaf to turn yellow. Since nutrients would have to be withdrawn from the leaves via the phloem, the pods do not induce yellowing by pulling nutrients out of the leaf and must be able to exert their influence via the xylem.     

Association of red coloration with senescence of sugar maple leaves in autumn

We evaluated the association of red coloration with senescence in sugar maple (Acer saccharum Marsh.) leaves by assessing differences in leaf retention strength and the progression of the abscission layer through the vascular bundle of green, yellow, and red leaves of 14 mature open-grown trees in October 2002. Computer image analysis confirmed visual categorization of leaves as predominantly green, yellow or red, and chemical quantification of leaf pigment concentrations verified that leaf color reflected underlying differences in leaf biochemistry. Significantly lower chlorophyll concentrations within red and yellow leaves indicated that senescence was more advanced in leaves from these color categories relative to green leaves. Among leaf types, only red leaves contained high concentrations of anthocyanins. There were significant differences in leaf retention capacity among color categories, with the petioles of green leaves being the most firmly attached to twigs, followed by red and then yellow leaves. Microscopic analysis indicated that yellow leaves had the most advanced extension of the abscission layer through the vasculature, with green and red leaves having significantly less abscission layer progression than yellow. A more limited progression of the abscission layer through vascular bundles may be evidence of delayed leaf senescence that could extend resorption of mobile leaf constituents. Together, results from this study suggest an association between leaf anthocyanin content and functional delays in senescence.     

Simple luminosity normalization of greenness, yellowness and redness/greenness for comparison of leaf spectral profiles in multi-temporally acquired remote sensing images

Observation of leaf colour (spectral profiles) through remote sensing is an effective method of identifying the spatial distribution patterns of abnormalities in leaf colour, which enables appropriate plant management measures to be taken. However, because the brightness of remote sensing images varies with acquisition time, in the observation of leaf spectral profiles in multi-temporally acquired remote sensing images, changes in brightness must be taken into account. This study identified a simple luminosity normalization technique that enables leaf colours to be compared in remote sensing images over time. The intensity values of green and yellow (green+red) exhibited strong linear relationships with luminosity (R2 greater than 0.926) when various invariant rooftops in Bangkok or Tokyo were spectralprofiled using remote sensing images acquired at different time points. The values of the coefficient and constant or the coefficient of the formulae describing the intensity of green or yellow were comparable among the single Bangkok site and the two Tokyo sites, indicating the technique's general applicability. For single rooftops, the values of the coefficient of variation for green, yellow, and red/green were 16% or less (n=6-11), indicating an accuracy not less than those of well-established remote sensing measures such as the normalized difference vegetation index. After obtaining the above linear relationships, raw intensity values were normalized and a temporal comparison of the spectral profiles of the canopies of evergreen and deciduous tree species in Tokyo was made to highlight the changes in the canopies' spectral profiles. Future aspects of this technique are discussed herein.     

罗汉果疱叶丛枝病的病原及其在某些寄主上的症状反应

葫芦科的罗汉果(Siraitia grosvenori(Swingle)Jeffrey)感染一种系统性的黄化型病害。 通过两次超薄切片的电镜观察:在病茎和病叶脉的韧皮组织中的薄壁细胞和伴胞中,发现有类菌质体(MLO),同时在重复的电镜观察中,在上述组织的薄壁细胞内,还发现风轮状的病毒(Virus)内含体。在健株的上述组织中。没有发现MLO和风轮状内含体。 在用三种不同接种方法(媒介昆虫、叶擦和汁液重创摩擦)进行的生物学测定的结果表明:罗汉果疱叶丛枝病有广泛的寄生范围,其症状特点:在常用的鉴定病毒的常规寄主上,最终均呈现黄化症状,没有系统性或局部性的斑点反应。其中在葫芦科的某些植物上的症状,基本上与罗汉果的症状相近似:初期明脉或脉间褪绿,继之出现疱状斑驳叶,叶片扭卷畸形,后期黄化。在茄科的供试植物上,症状特点:初期多为脉间褪绿,随之株形矮化,叶片变小。豆科上的症状:既有明脉,也有脉间褪绿,叶片呈现细长扭曲,变小并丛生。十字花科的症状特点是褪绿斑驳叶,株形矮化,叶片变小。     

Natural developmental variations in leaf and plant senescence in Arabidopsis thaliana

Leaf senescence is a developmentally regulated process that contributes to nutrient redistribution during reproductive growth and finally leads to tissue death. Manipulating leaf senescence through breeding or genetic engineering may help to improve important agronomic traits, such as crop yield and the storage life of harvested organs. Here, we studied natural variations in the regulation of plant senescence among 16 Arabidopsis thaliana accessions. Chlorophyll content and the proportion of yellow leaves were used as indicator parameters to determine leaf and plant senescence respectively. Our study indicated significant genotype effects on the onset and development of senescence. We selected three late- and five early-senescence accessions for further physiological studies. The relationship between leaf and plant senescence was accession-dependent. There was a significant correlation between plant senescence and the total number of leaves, siliques and plant bolting age. We monitored expression of two senescence marker genes, SAG12 and WRKY53 , to evaluate progression of senescence. Our data revealed that chlorophyll content does not fully reflect leaf age, because even fully green leaves had already commenced senescence at the molecular level. Integrating senescence parameters, such as the proportion of senescent leaves, at the whole plant level provided a better indication of the molecular status of the plant than single leaf senescence parameters.