Biyan Zhou

Hydrogen peroxide and nitric oxide promote reproductive growth in Litchi chinensis

Vegetative growth and reproductive growth strongly competes with each other during panicle development in litchi (Litchi chinensis Sonn.). We herein investigated the roles of hydrogen peroxide and nitric oxide in the competition between growth of rudimentary leaves and panicle development. The results show that the chilling-induced flowering increased H2O2 and NO contents in the mixed buds. Treatments with sodium nitroprusside (SNP), the NO donor, and methyl viologen dichloride hydrate (MV), the superoxide generator, increased NO and H2O2 contents in the mixed buds. MV and SNP treatments promoted abscission of rudimentary leaves and encouraged panicle development before or at the stage of panicle emergence. The nitric oxide synthase inhibitor Nω-nitro-L-arginine methyl ester (L-NAME) and the H2O2 trapper dimethylthiourea (DMTU) inhibited a chilling-induced flowering. SNP promoted the expression of litchi LEAFY homolog (LcLFY). These promotive effects were suppressed by the NO scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl3-oxide (PTIO) and the H2O2 trapper, DMTU. The results suggest that H2O2 and NO promote reproductive growth by inhibiting the growth of rudimentary leaves as well as by promoting the expression of the flower related gene, LcLFY.

Identification of nitric oxide responsive genes in the floral buds of Litchi chinensis

Litchi (Litchi chinensis Sonn.) is an important tropical and subtropical evergreen woody fruit tree, and it has been shown that nitric oxide (NO) could promote litchi flowering. NO responsive genes of litchi (cv. Nuomici) primordia were identified through a suppression subtractive hybridization (SSH) library screen. We obtained 1 563 expressed sequences tags (ESTs) that were enriched in the NO treated inflorescence primordia. We then used a reverse Northern analysis to identify 728 true NO responsive ESTs, the sequences of which have been further analyzed. They represent 70 litchi unique genes that could be classified into 9 categories: 14 % of them were involved in transport facilitation, 7 % in transcription regulation, 9 % in stress response, 7 % in sugar metabolism, 9 % in secondary metabolism, 10 % in intracellular signalling, and 44 % in other metabolism, whereas 11 % were genes with unknown functions, and 7 % were genes with no hit found. Next, we performed a real-time quantitative polymerase chain reaction (RT-qPCR) to determine the expression of selected candidate genes during a time-course of NO treatment and of normal floral tissue development.

Genome encode analyses reveal the basis of convergent evolution of fleshy fruit ripening

Fleshy fruits using ethylene to regulate ripening have developed multiple times in the history of angiosperms, presenting a clear case of convergent evolution whose molecular basis remains largely unknown. Analysis of the fruitENCODE data consisting of 361 transcriptome, 71 accessible chromatin, 147 histone and 45 DNA methylation profiles reveals three types of transcriptional feedback circuits controlling ethylene-dependent fruit ripening. These circuits are evolved from senescence or floral organ identity pathways in the ancestral angiosperms either by neofunctionalisation or repurposing pre-existing genes. The epigenome, H3K27me3 in particular, has played a conserved role in restricting ripening genes and their orthologues in dry and ethylene-independent fleshy fruits. Our findings suggest that evolution of ripening is constrained by limited hormone molecules and genetic and epigenetic materials, and whole-genome duplications have provided opportunities for plants to successfully circumvent these limitations.An analysis of the fruitENCODE data consisting of multiple transcriptome, accessible chromatin, histone and DNA methylation profiles from 11 fleshy fruits reveals three types of transcriptional feedback circuits controlling fruit ripening.

RNA-seq analysis of apical meristem reveals integrative regulatory network of ROS and chilling potentially related to flowering in Litchi chinensis

Litchi is an important woody fruit tree. Floral initiation in litchi is triggered by low temperatures. However, defective flowering is a major challenge for litchi production in times of climate change and global warming. Previous studies have shown that the reactive oxygen species (ROS) generated by methyl viologen dichloride hydrate (MV) promotes flowering. In this study, potted trees were transferred to growth chambers for low-temperature (LT), medium-temperature (MT), and high-temperature (HT) treatments. Trees at MT were subjected to ROS treatment to promote flowering, and those at LT were induced to flower. RNA-sequencing was applied to obtain a global transcriptome of the apical meristem and reveal potential gene networks controlling the transformation from vegetative meristems (VM) into inflorescence meristems (IM). We assembled 73,117 unigenes with a mean size of 790 bp and 11741 unigenes were identified as both chilling and ROS responsive genes (CRRGs), of which 48 were identified as flowering-related CRRGs, 59 were plant hormone signal transduction CRRGs, and 146 were plant hormone biosynthesis-related CRRGs. Genes co-expression network analysis indicated inner relationships, suggesting that ROS and chilling promotes the VM to IM transition through a regulatory gene network of transcription factors, hormones, and flowering regulators.

LcMCII-1 is involved in the ROS-dependent senescence of the rudimentary leaves of Litchi chinensis

Key messageLcMCII-1is a type II metacaspase. Over-expression ofLcMCII-1inArabidopsispromoted ROS-dependent and natural senescence. Virus-inducedLcMCII-1silencing delayed the ROS-dependent senescence of the rudimentary leaves ofLitchi chinensis.AbstractLitchi is an evergreen woody fruit tree that is widely cultivated in subtropical and tropical regions. Its floral buds are mixed with axillary or apical panicle primordia, leaf primordia and rudimentary leaves. A low spring temperature is vital for litchi production as it promotes the abscission of the rudimentary leaves, which could otherwise prevent panicle development. Hence, climate change could present additional challenges for litchi production. We previously reported that reactive oxygen species (ROS) can substitute low-temperature treatment to induce the senescence of rudimentary leaves. We have now identified from RNA-Seq data a litchi type II metacaspase gene, LcMCII-1, that is responsive to ROS. Silencing LcMCII-1 by virus-induced gene silencing delayed ROS-dependent senescence. The ectopic over-expression of LcMCII-1 in transgenic Arabidopsis promoted ROS-dependent and natural senescence. Consistently, the transient expression of LcMCII-1 in tobacco leaf by agroinfiltration resulted in leaf yellowing. Our findings demonstrate that LcMCII-1 is positively involved in the regulation of rudimentary leaf senescence in litchi and provide a new target for the future molecular breeding of new cultivars that can set fruit in warmer climates.

Low temperature-induced leaf senescence and the expression of senescence-related genes in the panicles of Litchi chinensis

Litchi is one of the most important subtropical evergreen fruit trees in Southern Asia. Litchi floral buds are a mix of axillary or apical panicle primordia, leaf primordia, and rudimentary leaves. Under usual winter and early spring conditions, the axillary panicle primordia prevail, and the rudimentary leaves abscise when low temperatures reach a certain threshold. The floral buds ultimately develop into pure panicles. Understanding the regulatory mechanism of rudimentary leaf senescence is of great importance for litchi flowering. In this study, litchi potted trees at the floral differentiation stage were treated with low and high temperatures in order to induce senescence or development of leaves. The microstructure of the petiole base of the rudimentary leaves was determined. The results show several layers of flattened cells forming in the abscission zone of the rudimentary leaves that were treated with low temperatures as well as an obvious boundary regarded as the abscission layer zone. We also determined the gene expression in the leaves with different developmental fate. The results show that the LcRboh, LcMC-1-like, and LcPirin genes were significantly induced in the rudimentary leaves treated with low temperatures, and the expression increased with the proceeding of senescence. The expression of the genes encoding class Ι β-1,3-glucanase and β-xylosidase also increased with the senescence, suggesting their possible involvement in the low temperature-induced senescence of the rudimentary leaves.

Reactive oxygen species and nitric oxide induce senescence of rudimentary leaves and the expression profiles of the related genes in Litchi chinensis

Litchi is one of the most important subtropical evergreen fruit trees in southern Asia. Previous studies indicated that high-temperature conditions encourage growth of rudimentary leaves in panicles and suppress flowering. We have demonstrated that methyl viologen dichloride hydrate (MV) and sodium nitroprusside (SNP) promoted flowering in litchi partially by inhibiting the growth of rudimentary leaves via reactive oxygen species (ROS) and nitric oxide (NO). In the present study, we examined the microstructure and ultrastructure, programmed cell death (PCD) ratio, nuclei morphology of the rudimentary leaves, and the expression of senescence-related genes after the treatment with ROS or NO. The results showed that chromatins of the ROS- or NO-treated cells in the rudimentary leaves were condensed. Fusion of the cytoplasm-digesting vesicles with the vacuole and degradation of cytoplasm forming scattered debris were found in those of the treated cells. Treatment with ROS or NO increased the cell PCD ratio. Morphology of the nuclei stained by propidium iodide (PI) showed that nuclei shape became irregular after the ROS or NO treatment. Further, the expression levels of LcRboh, LcMC-1-like, and LcPirin were higher in the ROS- and NO-treated rudimentary leaves than those in the control ones, suggesting that these genes may be involved in the ROS and NO-induced senescence and abscission of the rudimentary leaves in litchi. Our results suggested that ROS and NO play an important role in inducing the senescence of the rudimentary leaves, and ROS- and NO-induced PCD may be involved in the regulation of the rudimentary leaf growth in litchi.Climate change: tricking the treesWarmer winter temperatures suppress flowering of litchi trees, threatening fruit production, but researchers have found another way to trigger litchi trees to reproduce. In litchi, spring buds contain both flower and leaf primordia, and cold cues the plants to shed the young leaves, allowing the flower primordia to develop. With recent warmer winters, the leaves are crowding out the flowers. Biyan Zhou at South China Agricultural University investigated whether spraying with compounds that make the trees produce stress-signaling molecules, such as nitrous oxide, would mimic the effects of a cold winter. Following treatment, the trees shed their rudimentary leaves and flowers developed. Microscopic examination of buds showed that the leaf shedding process looked similar to cold-triggered shedding. This study offers a method to induce litchis to flower and fruit, even in a changing climate.