Transcription factor LBD16 targets cell wall modification/ion transport genes in peach lateral root formation.

LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2-LIKEs (LBDs/ASLs) are plant-specific transcription factors that function downstream of auxin-regulated lateral root (LR) formation. Our previous research found that PpLBD16 positively regulates peach (Prunus persica) LR formation. However, the downstream regulatory network and target genes of PpLBD16 are still largely unknown. Here, we constructed a PpLBD16 homologous overexpression line and a PpLBD16 silenced line. We found that overexpressing PpLBD16 promoted peach root initiation, while silencing PpLBD16 inhibited peach root formation. Through RNA sequencing (RNA-seq) analysis of roots from PpLBD16 overexpression and silenced lines, we discovered that genes positively regulated by PpLBD16 were closely related to cell wall synthesis and degradation, ion/substance transport, and ion binding and homeostasis. To further detect the binding motifs and potential target genes of PpLBD16, we performed DNA-affinity purification sequencing (DAP-seq) analysis in vitro. PpLBD16 preferentially bound to CCNGAAANNNNGG (MEME-1), [C/T]TTCT[C/T][T/C] (MEME-2), and GCGGCGG (ABR1) motifs. By combined analysis of RNA-seq and DAP-seq data, we screened candidate target genes for PpLBD16. We demonstrated that PpLBD16 bound and activated the cell wall modification-related genes EXPANSIN-B2 (PpEXPB2) and SUBTILISIN-LIKE PROTEASE 1.7 (PpSBT1.7), the ion transport-related gene CYCLIC NUCLEOTIDE-GATED ION CHANNEL 1 (PpCNGC1) and the polyphenol oxidase (PPO)-encoding gene PpPPO, thereby controlling peach root organogenesis and promoting LR formation. Moreover, our results displayed that PpLBD16 and its target genes are involved in peach LR primordia development. Overall, this work reveals the downstream regulatory network and target genes of PpLBD16, providing insights into the molecular network of LBD16-mediated LR development.

MdMYB305-MdbHLH33-MdMYB10 regulates sugar and anthocyanin balance in red-fleshed apple fruits.

Sugar and anthocyanin are important indicators of fruit quality, and understanding the mechanism underlying their accumulation is essential for breeding high-quality fruit. We identified an R2R3-MYB transcription factor MdMYB305 in the red-fleshed apple progeny, which was positively correlated with fruit sugar content but negatively correlated with anthocyanin content. Transient injection, stable expression [overexpressing and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)], and heterologous transformation of tomato confirmed that MdMYB305 promotes the accumulation of sugar and inhibits the synthesis of anthocyanin. A series of molecular experiments (such as electrophoretic mobility shift and luciferase assays) confirmed that MdMYB305 combines with sugar-related genes (MdCWI1/MdVGT3/MdTMT2) and anthocyanin-related genes (MdF3H/MdDFR/MdUFGT), promoting and inhibiting their activities, and finally regulating the sugar and anthocyanin content of fruits. In addition, the study also found that MdMYB305 competes with MdMYB10 for the MdbHLH33 binding site to balance sugar and anthocyanin accumulation in the fruits, which provides a reference value for exploring more functions of the MYB-bHLH-MYB complex and the balance relationship between sugar and anthocyanin in the future.

Abscisic acid and regulation of the sugar transporter gene MdSWEET9b promote apple sugar accumulation.

Enhancing fruit sugar contents, especially for high-flavonoid apples with a sour taste, is one of the main goals of horticultural crop breeders. This study analyzed sugar accumulation and the underlying mechanisms in the F2 progenies of a hybridization between the high-sugar apple (Malus × domestica) variety "Gala" and high-flavonoid apple germplasm "CSR6R6". We revealed that MdSWEET9b (sugars will eventually be exported transporter) helps mediate sugar accumulation in fruits. Functional characterization of MdSWEET9b in yeast mutants lacking sugar transport as well as in overexpressing and CRISPR/Cas9 knockdown apple calli revealed MdSWEET9b could transport sucrose specifically, ultimately promoting normal yeast growth and accumulation of total sugar contents. Moreover, MdWRKY9 bound to the MdSWEET9b promoter and regulated its activity, which responded to abscisic acid (ABA) signaling. Furthermore, MdWRKY9 interacted with MdbZIP23 (basic leucine zipper) and MdbZIP46, key ABA signal transducers, at the protein and DNA levels to enhance its regulatory effect on MdSWEET9b expression, thereby influencing sugar accumulation. Based on the contents of ABA in lines with differing sugar contents and the effects of ABA treatments on fruits and calli, we revealed ABA as one of the main factors responsible for the diversity in apple fruit sugar content. The results of this study have clarified how MdSWEET9b influences fruit sugar accumulation, while also further elucidating the regulatory effects of the ABA-signaling network on fruit sugar accumulation. This work provides a basis for future explorations of the crosstalk between hormone and sugar metabolism pathways.

Genome-wide identification of nucleotide-binding domain leucine-rich repeat (NLR) genes and their association with green peach aphid (Myzus persicae) resistance in peach.

Resistance genes (R genes) are a class of genes that are immune to a wide range of diseases and pests. In planta, NLR genes are essential components of the innate immune system. Currently, genes belonging to NLR family have been found in a number of plant species, but little is known in peach. Here, 286 NLR genes were identified on peach genome by using their homologous genes in Arabidopsis thaliana as queries. These 286 NLR genes contained at least one NBS domain and LRR domain. Phylogenetic and N-terminal domain analysis showed that these NLRs could be separated into four subfamilies (I-IV) and their promoters contained many cis-elements in response to defense and phytohormones. In addition, transcriptome analysis showed that 22 NLR genes were up-regulated after infected by Green Peach Aphid (GPA), and showed different expression patterns. This study clarified the NLR gene family and their potential functions in aphid resistance process. The candidate NLR genes might be useful in illustrating the mechanism of aphid resistance in peach.

Genome-wide identification, bioinformatics and expression analysis of HD-Zip gene family in peach.

BACKGROUND: HD-Zips (Homeodomain-Leucine Zippers) are a class of plant-specific transcription factors that play multiple roles in plant growth and development. Although some functions of HD-Zip transcription factor have been reported in several plants, it has not been comprehensively studied in peach, especially during adventitious root formation of peach cuttings. RESULTS: In this study, 23 HD-Zip genes distributed on 6 chromosomes were identified from the peach (Prunus persica) genome, and named PpHDZ01-23 according to their positions on the chromosomes. These 23 PpHDZ transcription factors all contained a homeomorphism box domain and a leucine zipper domain, were divided into 4 subfamilies(I-IV) according to the evolutionary analysis, and their promoters contained many different cis-acting elements. Spatio-temporal expression pattern showed that these genes were expressed in many tissues with different levels, and they had distinct expression pattern during adventitious root formation and development. CONCLUSION: Our results showed the roles of PpHDZs on root formation, which is helpful to better understand the classification and function of peach HD-Zip genes.

Silicon enhances the drought resistance of peach seedlings by regulating hormone, amino acid, and sugar metabolism.

BACKGROUND: Drought is one of the main concerns worldwide and restricts the development of agriculture. Silicon improves the drought resistance of plants, but the underlying mechanism remains unclear. RESULTS: We sequenced the transcriptomes of both control and silicon-treated peach seedlings under drought stress to identify genes or gene networks that could be managed to increase the drought tolerance of peach seedlings. Peach (Prunus persica) seedlings were used to analyse the effects of silicon on plant growth and physiological indexes related to drought resistance under drought stress. The results showed that silicon addition improved the water use efficiency, antioxidant capacity, and net photosynthetic rate, inhibition of stomatal closure, promoted the development of roots, and further regulated the synthesis of hormones, amino acids and sugars in peach seedlings. A comparative transcriptome analysis identified a total of 2275 genes that respond to silicon under drought stress. These genes were mainly involved in ion transport, hormone and signal transduction, biosynthetic and metabolic processes, stress and defence responses and other processes. We analysed the effects of silicon on the modulation of stress-related hormonal crosstalk and amino acid and sugar metabolism. The results showed that silicon promotes zeatin, gibberellin, and auxin biosynthesis, inhibits the synthesis of abscisic acid, then promote lateral root development and inhibit stomatal closure, and regulates the signal transduction of auxin, cytokinin, gibberellin and salicylic acid. Silicon also regulates the metabolism of various amino acids and promotes the accumulation of sucrose and glucose to improve drought resistance of peach seedlings. CONCLUSIONS: Silicon enhanced the drought resistance of peach seedlings by regulating stress-related hormone synthesis and signal transduction, and regulating amino acid and sugar metabolism.

Strawberry FaSnRK1α Regulates Anaerobic Respiratory Metabolism under Waterlogging.

Sucrose nonfermenting-1-related protein kinase 1 (SnRK1) is a central integrator of plant stress and energy starvation signalling pathways. We found that the FaSnRK1α-overexpression (OE) roots had a higher respiratory rate and tolerance to waterlogging than the FaSnRK1α-RNAi roots, suggesting that FaSnRK1α plays a positive role in the regulation of anaerobic respiration under waterlogging. FaSnRK1α upregulated the activity of anaerobic respiration-related enzymes including hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH) and lactate dehydrogenase (LDH). FaSnRK1α also enhanced the ability to quench reactive oxygen species (ROS) by increasing antioxidant enzyme activities. We sequenced the transcriptomes of the roots of both wild-type (WT) and FaSnRK1α-RNAi plants, and the differentially expressed genes (DEGs) were clearly enriched in the defence response, response to biotic stimuli, and cellular carbohydrate metabolic process. In addition, 42 genes involved in glycolysis and 30 genes involved in pyruvate metabolism were significantly regulated in FaSnRK1α-RNAi roots. We analysed the transcript levels of two anoxia-related genes and three ERFVIIs, and the results showed that FaADH1, FaPDC1, FaHRE2 and FaRAP2.12 were upregulated in response to FaSnRK1α, indicating that FaSnRK1α may be involved in the ethylene signalling pathway to improve waterlogging tolerance. In conclusion, FaSnRK1α increases the expression of ERFVIIs and further activates anoxia response genes, thereby enhancing anaerobic respiration metabolism in response to low-oxygen conditions during waterlogging.

Overexpression of PpSnRK1α in Tomato Increased Autophagy Activity under Low Nutrient Stress.

Plants suffer from a variety of environmental stresses during their growth and development. The evolutionarily conserved sucrose nonfermenting kinase 1-related protein kinase 1 (SnRK1) plays a central role in the regulation of energy homeostasis in response to stresses. In plant cells, autophagy is a degradation process occurring during development or under stress, such as nutrient starvation. In recent years, SnRK1 signaling has been reported to be an upstream activator of autophagy. However, these studies all focused on the regulatory effect of SnRK1 on TOR signaling and the autophagy-related gene 1 (ATG1) complex. In this study, overexpression of the gene encoding the Prunus persica SnRK1 α subunit (PpSnRK1α) in tomato improved the photosynthetic rates and enhanced the resistance to low nutrient stress (LNS). Overexpression of PpSnRK1α increased autophagy activity and upregulated the expression of seven autophagy-related genes (ATGs). The transcriptional levels of SlSnRK2 family genes were altered significantly by PpSnRK1α, signifying that PpSnRK1α may be involved in the ABA signaling pathway. Further analysis showed that PpSnRK1α not only activated autophagy by inhibiting target of rapamycin (TOR) signaling but also enhanced ABA-induced autophagy. This indicates that PpSnRK1α regulates the photosynthetic rate and induces autophagy, and then responds to low nutrient stress.

Phosphatidylcholine Enhances Homeostasis in Peach Seedling Cell Membrane and Increases Its Salt Stress Tolerance by Phosphatidic Acid.

Salt stress is a major adverse abiotic factor seriously affecting fruit tree growth and development. It ultimately lowers fruit quality and reduces yield. Phosphatidylcholine (PC) is an important cell membrane component that is critical for cell structure and membrane stability maintenance. In this study, we found that the addition of external PC sources significantly increased the tolerance of one-year-old peach trees, Prunus persica (L.) Batsch., to salt stress and attenuated their damage. The effect of exogenous application of 200 mg/L PC exerted the most significant positive effect. Its use caused seedling leaf stomatal opening, contributing to normal gas exchange. Moreover, beneficial effects were exerted also to the root system, which grew normally under salt stress. Meanwhile, phospholipase D activity in the cell was promoted. The production of phosphatidic acid (PA) was enhanced by increased decomposition of phospholipids; PA serves as a secondary messenger involved in plant biological process regulation and the reduction in the reactive oxygen species- and peroxide-induced damage caused by salt stress. The possible mechanism of action is via promoted plant osmotic regulation and tolerance to salt stress, reducing salt stress-induced injury to plants.

Comparative Transcriptome Analysis of Two Peach Rootstocks Uncovers the Effect of Gene Differential Expression on Nitrogen Use Efficiency.

Nitrogen is an important nutrient element that limits plant growth and yield formation, but excessive nitrogen has negative effects on plants and the environment. It is important to reveal the molecular mechanism of high NUE (nitrogen use efficiency) for breeding peach rootstock and variety with high NUE. In this study, two peach rootstocks, Shannong-1 (S) and Maotao (M), with different NUE were used as materials and treated with 0.1 mM KNO3 for transcriptome sequencing together with the control group. From the results of comparison between groups, we found that the two rootstocks had different responses to KNO3, and 2151 (KCL_S vs. KCL_M), 327 (KNO3_S vs. KCL_S), 2200 (KNO3_S vs. KNO3_M) and 146 (KNO3_M vs. KCL_M) differentially expressed genes (DEGs) were identified, respectively, which included multiple transcription factor families. These DEGs were enriched in many biological processes and signal transduction pathways, including nitrogen metabolism and plant hormone signal transduction. The function of PpNRT2.1, which showed up-regulated expression under KNO3 treatment, was verified by heterologous expression in Arabidopsis. The plant height, SPAD (soil and plant analyzer development) of leaf and primary root length of the transgenic plants were increased compared with those of WT, indicating the roles of PpNRT2.1 in nitrogen metabolism. The study uncovered for the first time the different molecular regulatory pathways involved in nitrogen metabolism between two peach rootstocks and provided gene reserve for studying the molecular mechanism of nitrogen metabolism and theoretical basis for screening peach rootstock or variety with high NUE.

PpSnRK1α overexpression alters the response to light and affects photosynthesis and carbon metabolism in tomato.

Sucrose nonfermentation 1 (SNF1) related kinase 1 (SnRK1) is a central energy sensor kinase in plants and a key switch regulating carbon and nitrogen metabolism. Fruit quality depends on leaf photosynthetic efficiency and carbohydrate accumulation, but the role of peach (Prunus persica) SnRK1 α subunit (PpSnRK1α) in regulating leaf carbon metabolism and the light signal response remains unclear. We studied the carbon metabolism of tomato leaves overexpressing PpSnRK1α and the responses of PpSnRK1α-overexpressing tomato leaves to light signals. Transcriptome, metabolome, and real-time quantitative polymerase chain reaction analyses revealed that uridine 5'-diphosphoglucose, glutamate, and glucose-6-phosphate accumulated in tomato leaves overexpressing PpSnRK1α. The expression of genes (e.g., GDH2, SuSy) encoding enzymes related to carbon metabolism (e.g., glutamate dehydrogenase (GDH2; EC: 1.4.1.3), sucrose synthase (SS; EC: 2.4.1.13)) and chlorophyllase (CLH) encoding chlorophyllase (EC: 3.1.1.14), which regulates photosynthetic pigments and photosynthesis, was significantly increased in PpSnRK1α-overexpressing plants. PpSnRK1α overexpression inhibited the growth of hypocotyls and primary roots in response to light. The chlorophyll content of the leaves was increased, the activity of SS and ADPG pyrophosphatase (AGPase; EC: 2.7.7.27) was increased, and photosynthesis was promoted in PpSnRK1α-overexpressing plants relative to wild-type plants. Under light stress, the net photosynthetic rate of plants was significantly higher in plants overexpressing PpSnRK1α than in wild-type plants. This indicates that PpSnRK1α promotes the accumulation of carbohydrates by regulating genes related to carbon metabolism, regulating genes related to chlorophyll synthesis, and then responding to light signals to increase the net photosynthetic rate of leaves.

Overexpression of PpSnRK1α in tomato enhanced salt tolerance by regulating ABA signaling pathway and reactive oxygen metabolism.

BACKGROUND: SNF-related Kinase 1 (SnRK1) is a key component of the cell signaling network. SnRK1 is known to respond to a wide variety of stresses, but its exact role in salt stress response and tolerance is still largely unknown. RESULTS: In this study, we reported that overexpression of the gene encoding the α subunit of Prunus persica SnRK1 (PpSnRK1α) in tomato could improve salt stress tolerance. The increase in salt stress tolerance in PpSnRK1α-overexpressing plants was found to correlate with increased PpSnRK1α expression level and SnRK1 kinase activity. And PpSnRK1α overexpression lines exhibited a lower level of leaf damage as well as increased proline content and reduced malondialdehyde (MDA) compared with wild-type (WT) lines under salt stress. Furthermore, PpSnRK1α enhanced reactive oxygen species (ROS) metabolism by increasing the expression level of antioxidase genes and antioxidant enzyme activities. We further sequenced the transcriptomes of the WT and three PpSnRK1α overexpression lines using RNA-seq and identified about 1000 PpSnRK1α-regulated genes, including many antioxidant enzymes, and these genes were clearly enriched in the MAPK signaling pathway (plant), plant-pathogen interactions and plant hormone signaling transduction and can respond to stimuli, metabolic processes, and biological regulation. Furthermore, we identified the transcriptional levels of several salt stress-responsive genes, SlPP2C37, SlPYL4, SlPYL8, SlNAC022, SlNAC042, and SlSnRK2 family were altered significantly by PpSnRK1α, signifying that SnRK1α may be involved in the ABA signaling pathway to improve tomato salt tolerance. Overall, these findings provided new evidence for the underlying mechanism of SnRK1α conferment in plant salt tolerance phenotypes. CONCLUSIONS: Our findings demonstrated that plant salt stress resistance can be affected by the regulation of the SnRK1α. Further molecular and genetic approaches will accelerate our knowledge of PpSnRK1α functions, and inform the genetic improvement of salt tolerance in tomato through genetic engineering and other related strategies.

The effect of auxin and strigolactone on ATP/ADP isopentenyltransferase expression and the regulation of apical dominance in peach.

KEY MESSAGE: We confirmed the roles of auxin, CK, and strigolactones in apical dominance in peach and established a model of plant hormonal control of apical dominance in peach. Auxin, cytokinin, and strigolactone play important roles in apical dominance. In this study, we analyzed the effect of auxin and strigolactone on the expression of ATP/ADP isopentenyltransferase (IPT) genes (key cytokinin biosynthesis genes) and the regulation of apical dominance in peach. After decapitation, the expression levels of PpIPT1, PpIPT3, and PpIPT5a in nodal stems sharply increased. This observation is consistent with the changes in tZ-type and iP-type cytokinin levels in nodal stems and axillary buds observed after treatment; these changes are required to promote the outgrowth of axillary buds in peach. These results suggest that ATP/ADP PpIPT genes in nodal stems are key genes for cytokinin biosynthesis, as they promote the outgrowth of axillary buds. We also found that auxin and strigolactone inhibited the outgrowth of axillary buds. After decapitation, IAA treatment inhibited the expression of ATP/ADP PpIPTs in nodal stems to impede the increase in cytokinin levels. By contrast, after GR24 (GR24 strigolactone) treatment, the expression of ATP/ADP IPT genes and cytokinin levels still increased markedly, but the rate of increase in gene expression was markedly lower than that observed after decapitation in the absence of IAA (indole-3-acetic acid) treatment. In addition, GR24 inhibited basipetal auxin transport at the nodes (by limiting the expression of PpPIN1a in nodal stems), thereby inhibiting ATP/ADP PpIPT expression in nodal stems. Therefore, strigolactone inhibits the outgrowth of axillary buds in peach only when terminal buds are present.

Lipid Metabolomic and Transcriptomic Analyses Reveal That Phosphatidylcholine Enhanced the Resistance of Peach Seedlings to Salt Stress through Phosphatidic Acid.

Soil salinity is a major conlinet limiting sustainable agricultural development in peach tree industry. In this study, lipid metabolomic pathway analysis indicated that phosphatidic acid is essential for root resistance to salt stress in peach seedlings. Through functional annotation analysis of differentially expressed genes in transcriptomics, we found that MAPK signaling pathway is closely related to peach tree resistance to salt stress, wherein PpMPK6 expression is significantly upregulated. Under salt conditions, the OE-PpMPK6 Arabidopsis thaliana (L.) Heynh. line showed higher resistance to salt stress than WT and KO-AtMPK6 lines. Furthermore, we found that the Na+ content in OE-PpMPK6 roots was significantly lower than that in WT and KO-AtMPK6 roots, indicating that phosphatidic acid combined with PpMPK6 activated the SOS1 (salt-overly-sensitive 1) protein to enhance Na+ efflux, thus alleviating the damage caused by NaCl in roots; these findings provide insight into the salt stress-associated transcriptional regulation.

Effects of Valine and Urea on Carbon and Nitrogen Accumulation and Lignin Content in Peach Trees.

Nitrogen availability and uptake levels can affect nutrient accumulation in plants. In this study, the effects of valine and urea supplementation on the growth of new shoots, lignin content, and carbon and the nitrogen metabolism of 'Ruiguang 39/peach' were investigated. Relative to fertilization with urea, the application of valine inhibited shoot longitudinal growth, reduced the number of secondary shoots in autumn, and increased the degree of shoot lignification. The application of valine also increased the protein level of sucrose synthase (SS) and sucrose phosphate synthase (SPS) in plant leaves, phloem, and xylem, thereby increasing the soluble sugar and starch content. It also resulted in an increase in nitrate reductase (NR), glutamine synthase (GS), and glutamate synthase (GOGAT) protein levels, with an increase in plant contents of ammonium nitrogen, nitrate nitrogen, and soluble proteins. Although urea application increased the protein level of carbon- and nitrogen-metabolizing enzymes, the increase in plant growth reduced the overall nutrient accumulation and lignin content per unit tree mass. In conclusion, the application of valine has a positive effect on increasing the accumulation of carbon and nitrogen nutrients in peach trees and increasing the lignin content.

Dynamic Monitoring of Nutrition Inputs and Fertility Evaluation during a Decade in the Main Peach-Producing Areas of Shandong Province, China.

The main peach-producing area in Shandong is an important peach fruit-producing area in China. Understanding the nutritional properties of the soil in peach orchards helps us to understand the evolution of soil properties and adjust management methods in a timely manner. This study focuses on 52 peach orchards in the main peach-producing area in Shandong as the research object. The spatiotemporal changes in soil traits and their influential factors were studied in depth, and the changes in soil fertility were effectively evaluated. The results showed that the input of nitrogen, phosphorus and potassium from organic fertilizer in 2021 was significantly higher than that in 2011, while the input of fertilizer in 2011 was significantly higher than that in 2021. Compared with traditional parks, both organic fertilizer inputs and chemical fertilizer inputs in demonstration parks showed a significant downwards trend. There was no significant change in pH values between 2011 and 2021. In 2021, the soil organic matter (SOM) contents of the 0-20 cm and 20-40 cm layers were 24.17 g·kg-1 and 23.38 g·kg-1, respectively, an increase of 29.3% and 78.47% over the values measured in 2011. Compared with 2011, the content of soil alkaloid nitrogen (AN) decreased significantly in 2021, and the contents of available phosphorus (AP) and available potassium (AK) in the soil increased significantly. According to the calculation results of the comprehensive fertility index (IFI) value, we found that in 2021, compared with 2011, the quality of soil fertility improved, most of which was at the medium and high levels. The research results show that the fertilizer-saving and synergistic approach in peach orchards in China significantly improved the soil nutrition. In the future, research on suitable comprehensive technologies should be strengthened in the management of peach orchards.

Physiological and Transcriptomic Analyses of the Effects of Exogenous Lauric Acid on Drought Resistance in Peach (Prunus persica (L.) Batsch).

Peach (Prunus persica (L.) Batsch) is a fruit tree of economic and nutritional importance, but it is very sensitive to drought stress, which affects its growth to a great extent. Lauric acid (LA) is a fatty acid produced in plants and associated with the response to abiotic stress, but the underlying mechanism remains unclear. In this study, physiological analysis showed that 50 ppm LA pretreatment under drought stress could alleviate the growth of peach seedlings. LA inhibits the degradation of photosynthetic pigments and the closing of pores under drought stress, increasing the photosynthetic rate. LA also reduces the content of O2-, H2O2, and MDA under drought stress; our results were confirmed by Evans Blue, nitroblue tetrazolium (NBT), and DAB(3,3-diaminobenzidine) staining experiments. It may be that, by directly removing reactive oxygen species (ROS) and improving enzyme activity, i.e., catalase (CAT) activity, peroxidase (POD) activity, superoxide dismutase (SOD) activity, and ascorbate peroxidase (APX) activity, the damage caused by reactive oxygen species to peach seedlings is reduced. Peach seedlings treated with LA showed a significant increase in osmoregulatory substances compared with those subjected to drought stress, thereby regulating osmoregulatory balance and reducing damage. RNA-Seq analysis identified 1876 DEGs (differentially expressed genes) in untreated and LA-pretreated plants under drought stress. In-depth analysis of these DEGs showed that, under drought stress, LA regulates the expression of genes related to plant-pathogen interaction, phenylpropanoid biosynthesis, the MAPK signaling pathway, cyanoamino acid metabolism, and sesquiterpenoid and triterpenoid biosynthesis. In addition, LA may activate the Ca2+ signaling pathway by increasing the expressions of CNGC, CAM/CML, and CPDK family genes, thereby improving the drought resistance of peaches. In summary, via physiological and transcriptome analyses, the mechanism of action of LA in drought resistance has been revealed. Our research results provide new insights into the molecular regulatory mechanism of the LA-mediated drought resistance of peach trees.

Waterlogging resistance and evaluation of physiological mechanism of three peach (Prunus persica) rootstocks.

Waterlogging occurs due to poor soil drainage or excessive rainfall. It is a serious abiotic stress factor that negatively affects crop growth. Waterlogging often causes plants to shed leaves, fruits, and, ultimately, to die. Peach (Prunus persica) trees are generally intolerant to waterlogging, and the primary peach rootstock used in Chinais "Maotao," which has very poor resistance to sensitivity. Therefore, waterlogging has become a restriction on the development of the peach industry in many regions. In this experiment, we tested the waterlogging resistance of "Maotao (Prunus persica (L.) Batsch)" (MT), "Shannong1 (GF677 × Cadaman)" (SN1), and "Mirabolano 29C (Prunus cerasifera)" (M29C) rootstocks. Using a simulated waterlogging method, the effects of waterlogging on the photosynthetic system, leaf pigments, osmotic adjustment, lipid membrane peroxidation, and antioxidant system of these three peach rootstocks were studied, and the changes of chlorophyll fluorescence parameters and fluorescence imaging were observed. The results showed that, with prolonged waterlogging, the photosynthetic pigment content and photosynthesis of the three peach rootstocks decreased rapidly, but the decomposition rate of SN1 and M29C chlorophyll was slower, and it still had high light energy absorption and energy transfer capabilities under waterlogging stress, which reduced the damage caused by waterlogging stress; under the stress of flooding, the osmoregulatory substances of the three rootstocks increased to varying degrees compared with normal conditions. At the same time, the enzyme activity of superoxide dismutase (SOD) activity, peroxidase (POD) activity, and catalase (CAT) activity in the leaves of the three rootstocks under flooding stress all increased and then decreased; during this period, malondialdehyde (MDA) continued to increase, and SN1 and M29C were significantly lower than MT; and chlorophyll fluorescence parameters, including the maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ΦPSII), photochemical quenching coefficient (qP), non-photochemical quenching (NPQ), and electron transfer rate (ETR) decreased significantly. The tolerance of SN1 and M29C to waterlogging was significantly better than that of MT rootstocks. The rootstock and grafted seedlings of SN1 have good waterlogging tolerance.

Genome-Wide Identification of Trehalose-6-phosphate Synthase (TPS) Gene Family Reveals the Potential Role in Carbohydrate Metabolism in Peach.

Trehalose-6-phosphate synthase (TPS) is essential for plant growth and development, linking trehalose-6-phosphate (T6P) to carbon metabolism. However, little is known about the TPS gene family in peaches and their potential roles in regulating carbohydrates in peach fruit. In this study, nine TPS genes were identified in the peach genome and named according to the homologous genes in Arabidopsis. Phylogenetic analysis showed that three subfamilies were identified, including TPSI, TPSII-1, and TPSII-2, which were also consistent with gene structure analysis. Considerable cis-elements were enriched in the promoters, including plant hormone-related elements. Tissue-specific analysis showed that these TPS genes were mainly expressed in leaves, stems, and fruit, showing different expression patterns for each gene. In addition, during fruit development, the content of trehalose-6-phosphate (T6P) was positively correlated with the expression of PpTPS7a and negatively with sucrose non-fermenting-1-related kinase 1 (SnRK1) activity. Transient overexpression and silencing of PpTPS7a in peach fruit validated its function in regulating T6P content and SnRK1 activity.

Molecular cloning and characterization of a nonsymbiotic hemoglobin gene (GLB1) from Malus hupehensis Rehd. with heterologous expression in tomato.

Nonsymbiotic hemoglobins (nsHbs) are involved in a variety of cellular processes in plants. Previous studies indicate that nsHb expression improves plant tolerance during waterlogging and hypoxia. In the present work, the nsHb class-1 coding sequence was cloned from Malus hupehensis Rehd. var. pinyiensis Jiang and subsequently named MhGLB1. The results elucidated the expressed characteristics and physiological effects of MhGLB1. The full-length cDNA contained a 477 bp open reading frame encoding a protein with a molecular mass of 17.8 KDa with 158 amino acids. Quantitative real-time PCR analysis showed that MhGLB1 expresses in roots, stems and leaves growing under normal and nitrate-induced conditions. Hypoxic stress induced accumulation of MhGLB1 within 12 h, and abscisic acid significantly induced expression of MhGLB1 in roots. The photosynthetic, transpiration and stomatal conductance rates of transgenic MhGLB1 tomato plants decreased more slowly than that of wild-type plants under waterlogging treatment. These results indicated that the MhGLB1 gene has an important role in hypoxia.