Genome-wide identification of the CONSTANS-LIKE(COL)family and mechanism of fruit senescence regulation by PpCOL8 in sand pear(Pyrus pyrifolia)

Pyrus pyrifolia Nakai‘Whangkeumbae'is a sand pear fruit with excellent nutritional quality and taste.However,the industrial development of pear fruit is significantly limited by its short shelf life.Salicylic acid(SA),a well-known phytohormone,can delay fruit senescence and improve shelf life.However,the mechanism by which SA regulates CONSTANS-LIKE genes(COLs)during fruit senescence and the role of COL genes in mediating fruit senescence in sand pear are poorly understood.In this study,22 COL genes were identified in sand pear,including four COLs(Pp COL8,Pp COL9a,Pp COL9b,and Pp COL14)identified via transcriptome analysis and 18 COLs through genome-wide analysis.These COL genes were divided into three subgroups according to the structural domains of the COL protein.Pp COL8,with two B-box motifs and one CCT domain,belonged to the first subgroup.In contrast,the other three Pp COLs,Pp COL9a,Pp COL9b,and Pp COL14,with similar conserved protein domains and gene structures,were assigned to the third subgroup.The four COLs showed different expression patterns in pear tissues and were preferentially expressed at the early stage of fruit development.Moreover,the expression of Pp COL8 was inhibited by exogenous SA treatment,while SA up-regulated the expression of Pp COL9a and Pp COL9b.Interestingly,Pp COL8 interacts with Pp MADS,a MADS-box protein preferentially expressed in fruit,and SA up-regulated its expression.While the production of ethylene and the content of malondialdehyde(MDA)were increased in Pp COL8-overexpression sand pear fruit,the antioxidant enzyme(POD and SOD)activity and the expression of Pp POD1 and Pp SOD1 in the sand pear fruits were down-regulated,which showed that Pp COL8 promoted sand pear fruit senescence.In contrast,the corresponding changes were the opposite in Pp MADS-overexpression sand pear fruits,suggesting that Pp MADS delayed sand pear fruit senescence.The co-transformation of Pp COL8 and Pp MADS also delayed sand pear fruit senescence.The results of this study revealed that Pp COL8 can play a key role in pear fruit senescence by interacting with Pp MADS through the SA signaling pathway.

Insights into the dwarfing mechanism of pear(Pyrus betulaefolia) based on anatomical and structural analysis using X-ray scanning

The lack of a suitable rootstock to control scion growth has limited the development of high-density plantations in pear production, which is partly attributed to poor understanding of the dwarfing mechanism. In the present study, the rootstock of the dwarf-type pear (Pyrus betulaefolia)PY-9’ was identified and used as the material for anatomical analysis.PY-9’ grew to half the tree height of the normal cultivar Zhengdu’, along with fewer internodes and shorter length. Significant differences in growth rate betweenPY-9’ andZhengdu’ were detected at approximately 30 days after full bloom, which corresponded with the time of the greatest difference in water potential between the dwarf and normal cultivar.PY-9’ showed a higher photosynthetic rate thanZhengdu’. Anatomical analysis showed thatPY-9’ had higher area ratios of both phloem and xylem and more developed vascular tissues thanZhengdu’. The three-dimensional reconstructed skeleton of the xylem from X-ray computed tomography scanning revealed greater intervessel connectivity inZhengdu’ than inPY-9’, which could contribute to the more vigorous growth ofZhengdu’. This study thus provides the first comparison of the microstructural properties of xylem elements between a dwarfing-type and vigorous-type pear rootstock, providing new insights into the dwarfing mechanism in pear and facilitating breeding of dwarf pear rootstocks to increase crop productivity.

Integrative transcriptomic and metabolomic analyses reveal the flavonoid biosynthesis of Pyrus hopeiensis flowers under cold stress

Low temperature is among the most restrictive factors to limit the yield and distribution of pear. Pyrus hopeiensis is a valuable wild resource.PCA showed that P. hopeiensis had strong cold resistance. In this study, the mRNA and metabolome sequencing of P. hopeiensis flower organs exposed to different low temperatures were performed to identify changes of genes and metabolites in response to low-temperature stress. A total of 4 851 differentially expressed genes(DEGs) were identified. Trend analysis showed that these DEGs were significantly enriched in profiles 19, 18, 7, 14, 1, 4 and 11. And the KEGG enrichment analysis showed that the DEGs in profile 18 were significantly enriched in flavone and flavonol biosynthesis. Besides, the expressed trends as well as GO and KEGG functional enrichment analyses of DEGs under cold and freezing stress showed significantly difference. Analyses of flavonoid-related pathways indicated that flavonoid structural genes had undergone significant changes. Correlation analysis showed that b HLH and MYB TFs may affect flavonoid biosynthesis by regulating structural gene expression. And PhMYB308 and PhMYB330 were likely candidate repressors of flavonoid biosynthesis by binding to a specific site in bHLH proteins. In total, 92 differentially accumulated metabolites(DAMs) were identified in P. hopeiensis flowers including 12 flavonoids. WGCNA results showed that coral 1, pink and brown 4 modules were closely associated with flavonoids and 11 MYBs and 15 bHLHs among the three modules may activate or inhibit the expression of 23 structural genes of flavonoid biosynthesis. Taken together, the results of this study provided a theoretical basis for further exploration of the molecular mechanisms of flavonoid biosynthesis and cold resistance of P. hopeiensis flower organs and our findings laid a foundation for further molecular breeding in cold-resistant pear varieties.

The PcERF5 promotes anthocyanin biosynthesis in red-fleshed pear(Pyrus communis)through both activating and interacting with PcMYB transcription factors

As there is a strong interest in red-skinned pears,the molecular mechanism of anthocyanin regulation in red-skinned pears has been widely investigated;however,little is known about the molecular mechanism of anthocyanin regulation in red-fleshed pears due to limited availability of such germplasm,primarily found in European pears(Pyrus communis).In this study,based on transcriptomic analysis in red-fleshed and white-fleshed pears,we identified an ethylene response factor(ERF)from P.communis,PcERF5,of which expression level in fruit flesh was significantly correlated with anthocyanin content.We then verified the function of PcERF5 in regulating anthocyanin accumulation by genetic transformation in both pear skin and apple calli.PcERF5 regulated anthocyanin biosynthesis by different regulatory pathways.On the one hand,PcERF5 can activate the transcription of flavonoid biosynthetic genes(PcDFR,PcANS and PcUFGT)and two key transcription factors encoding genes PcMYB10 and PcMYB114.On the other hand,PcERF5 interacted with PcMYB10 to form the ERF5-MYB10 protein complex that enhanced the transcriptional activation of PcERF5 on its target genes.Our results suggested that PcERF5 functioned as a transcriptional activator in regulating anthocyanin biosynthesis,which provides new insights into the regulatory mechanism of anthocyanin biosynthesis.This new knowledge will provide guidance for molecular breeding of red-fleshed pear.

Identification and evolutionary characterization of SFBB genes in‘Yali’and its spontaneous self-compatible mutant‘Jinzhui’(Pyrus bretschneideri)

Pears carry a gametophytic self-incompatibility(SI)system.In this system,S-RNase is the SI pistil determinant,and S-locus F-box brothers(SFBBs)are candidate pollen determinants.However,compared with apple,fewer SFBB genes were identified from pear,possibly caused by the lack of economic and effective methods.Here,we used transcriptome sequencing on‘Yali’(Pyrus bretschneideri)to obtain sequence fragments of SFBB genes and then used polymerase chain reaction(PCR)to amplify the whole sequence of SFBB genes.Twenty-seven SFBB genes,including22 full-length and five nonfull-length SFBB genes,were identified in‘Yali’(P.bretschneideri).SFBBs linkage analysis by PCR-enzyme-linked immunoassay(ELISA)showed that 12 SFBB genes belong to the S21 locus,and 15 SFBB genes belong to the S34 locus.Phylogenetic analysis showed that SFBB genes from Pyrus were divided into 26 types,more than the original eight types.The intrahaplotypic divergence of SFBBs is high and comparable to the allelic diversity of S-RNase,which is consistent with a nonself-recognition SI system.In addition,the expression level of PbrSFBBs in‘Jinzhui’,the only known haploid pollen of a self-compatible mutant,was mostly approximately two times higher than in‘Yali’,which may be the reason for the self-compatible mutant.

Adaptive responses of Acer ginnala, Pyrus ussuriensis and Prunus davidiana seedlings to soil moisture stress

One-year-old seedlings of Amur maple (Acer ginnala Maxim), Ussurian pear (Pyrus ussuriensis Maxim) and David peach (Prunus davidiana Carr) were planted in pots in greenhouse and treated with four different soil moisture contents (75.0%, 61.1%, 46.4% and 35.4%). The results showed that net photosynthesis rate (NPR), transpiration rate (TR) and stomatal conductance (Sc) of seedlings of the three species decreased with the decease of soil moisture content, and Amur maple seedlings had the greatest change in those physiological indices, followed by Ussurian pear, David peach. Amur maple and Ussurian pear seedlings also presented a decrease tendency in water use efficiency (WUE) under lower soil moisture content, whereas this was reversed for David peach. Under water stress the biomass allocation to seedling root had a significant increase for all the experimental species. As to root/shoot ratio, Amur maple seedlings had the biggest increase, while David peach had the smallest increase. The leaf plasticity of Amur maple seedlings was greater, the leaf size and total leaf area decreased significantly as the stress was intensified. No significant change of leaf size and total leaf area was found in seedlings of Ussurian pear and David peach. It was concluded that Amur maple was more tolerant to soil moisture stress in comparison with David peach and Ussurian pear.

Identification of Self-Incompatibility Genotypes in Some Sand Pears (Pyrus pyrifolia Nakai) by PCR-RFLP Analysis

The identification of self-incompatibility genotype （S-genotype） will be useful for selection of pollinizers and design of crossing in cultivar improvement of sand pear. This paper reported the identification of self-incompatibility genotypes of seven Chinese and two Japanese sand pear cultivars using PCR-RFLP analysis and S-RNase sequencing. The Sgenotypes of these cultivars were determined as follows： Huali 1 S1S3, Shounan S1S3, Xizilti S1S4, Qingxiang S3S7, Sanhua S2S7, Huangmi （Imamuranatsu） S1S6, Huali 2 S3S4, Baozhuli S7S33, Cangxixueli S5S15. S-RNase alleles （S1 to S9） in sand pear could be identified effectively by PCR-RFLP analysis.

Relationship Between Anthocyanin Biosynthesis and Related Enzymes Activity in Pyrus pyrifolia Mantianhong and Its Bud Sports Aoguan

The aim of this article is to study the relationship between biosynthesis of anthocyanin and activities of phenylalanine ammonia lyase （PAL）, chalcone ismoerase （CHI） enzymes in Pyrus pyrifolia. Changes in the level of anthocyanin and the activities of enzymes of anthocyanin biosynthesis including PAL, CHI were studied in the pericarp of Pyrus pyrifolia Aoguan and Mantianhong during the period of pigment formation. Bagging treatment was also carried out to manipulate the synthesis of anthocyanin and the activities of related enzymes during the period of pigment formation. The results demonstrated that the level of anthocyanin of Aoguan was higher than that of Mantianhong. However, the content of anthocyanins has the similar changing trend in Aoguan and Mantianhong, highest anthocyanin concentrations of two varieties appeared in immature fruit and faded toward harvest. Meanwhile, similar changing trends of activities of PAL and CHI were also observed in both varieties. Aoguan has a lower activity of PAL than Mantianhong, whereas activity of CHI in Aoguan was higher than that in Mantianhong. Activity of PAL decreased during the period of pigment formation and was apparently not limiting to color development, whereas CHI activity increased at the same period and was closely related to the synthesis of anthocyanin. The results of bagging treatment showed that bagging treatment inhibited the activity of CHI, as well as the synthesis of anthocyanin, whereas debagging enhanced both the activity of CHI and synthesis of anthocyanin. The activity of CHI in debagging Aoguan pericarp was higher than the untreated Aoguan. However, effect of bagging treatment toward PAL activity was not obvious. Anthocyanin of bagging treated Aoguan decreased toward harvest. The content of anthocyanin of Pyruspyrifolia increased at the beginning of fruit coloration period and decreased toward fruit harvest. Activity of PAL was apparently not limiting to color development, whereas CHI activity was closely related to the synthesis of anthocyanin. Debagging enhanced both the activity of CHI and anthocyanin synthesis. Bagging treatment also proved that degradation of anthocyanin was not induced only by light. Light appeared to have two opposing effects in pears： it is required for anthocyanin synthesis and also for apparently increasing red color loss through increased degradation of anthocyanin.

Cloning and Analysis of Full-Length cDNA of PumNPR1 Gene from Pyrus ussuriensis Maxim

The purpose of this study is to find a new gene resource for the researches of molecular breeding of Rosaceae plants disease-resistance. Pyrus ussuriensis Maxim is used as a starting material to clone the full-length cDNA of NPR1（nonexpressor of pathogenesis- related genes 1） which is a key regulator in SA （salicylic acid）-mediated systemic acquired resistance （SAR） by homologous cloning and RACE techniques. The length of the cDNA sequence was 1 767 bp, the ORF was 1 761 bp, it coded 586 amino acids, pi=5.58, the relative molecular weight was 65.009 ku, contained 19 kinds of amino acids, and had full BTB/POZ and ANK domains. Compared the homology of NPR1 gene in GenBank database, the homology with Pyrus pyrifolia, Arabidopsis thaliana, Nicotiana tabacum, Lycopersicon esculentum, Oryza sativa, Helianthus annuus were 98%, 62%, 68%, 65%, 57%, 63%. The homology offunctional area were 99%, 78%, 82%, 79%, 74%, 77%. This NPR1 gene was considered as homologic gene of Pyrus ussuriensis Maxim and named PumNPR1.

Isolation and Characterization of a Cinnamoyl CoA Reductase Gene(CCR) in Pear(Pyrus pyrifolia)

Pear is a popular and commercially important fresh fruit, and its texture is related to the presence of sclereid formatted by parenchyma cell with lignification in vascular plants. Previous studies have demonstrated that content of lignin may be regulated by cinnamoyl CoA reductase(CCR) in various plants. However, the function of CCR in pears remains very limited. In the present study, we isolated a cDNA encoding CCR(PpCCR, GenBank accession No. KF999958) and its promoter(proPpCCR) from Whangkeumbae pear to investigate the function of CCR in lignin biosynthesis. PpCCR-GFP expressed in rice mesophyll protoplast demonstrated that PpCCR-GFP was localized in the cytoplasm, indicating that CCR may function in cytoplasm without localization signals. In transgenic plants carrying PpCCR, we observed higher lignin content compared with that in wild type plants, further suggesting that PpCCR can affect the lignin contents through regulating lignin biosynthesis in Arabidopsis thaliana. More studies in other plants are needed to confirm our conclusion.

Physiological Effects of Iron Deficiency on Pyrus pashia Buch-Ham

Pyrus pashia Buch-Ham, a wild specie was used to investigate the physiological effects of iron deficiency in culture solution. The result showed that Chla, Chlb, total chlorophyll content and photosynthesis rate(Pn) decreased sharply, and the decrease of Pn was prior to that of Chl content under the iron deficiency. The iron deficiency symptoms were visible when the iron concentration in culture medium was less than 25 μmol L-1. Peroxidase(POD) and catalase(CAT) activity in iron-deficient leaves declined significantly, and POD was more sensitive than CAT to Fe deficiency. However, the positive correlation between CAT activity and Chl content was more significant than that between POD activity and Chl content. The content of nutrient elements in Fe-deficient leaves, which changed irregularly, were higher than that in normal leaves. There were a most significant positive correlation between active Fe and Chl content, and between active Fe and Pn respectively. Therefore, active Fe could be useful physiological predicting index for diagnosis.

Advances in Origin,Evolution and Classification of Pyrus L.

China is not only one of the origin centers of Pyrus L.,but also the earliest birthplace of Pyrus L.in the world.This paper reviews the evolution of Pyrus L.from the aspects of leaf edge morphology,inflorescence and fruit type,and summarizes the research progress of classification and species distribution of Pyrus L.,which is of great significance for the protection,evaluation and utilization of germplasm resources.

PbrARF4 contributes to calyx shedding of fruitlets in ‘Dangshan Suli’ pear by partly regulating the expression of abscission genes

Fruitlet calyx shedding in pear plants is apparently regulated via numerous pathways that involve both environmental triggers and phytohormones cues such as auxin. In this study, we found at 10 days after full bloom (DAFB) higher levels of indoleacetic acid (IAA) and tryptophan (Trp) in calyx persistence fruitlet (CPF) than calyx shedding fruitlet (CSF) ofDanshan Suli’ pear (Pyrus bretschneideri Rhed.). Consisting with this, the activity of indolealdehyde oxidase (IAAIdO), which promotes IAA synthesis, was remarkably increased, and that of peroxidase(POD), which degrades IAA, dropped markedly in CPF but not in CSF. Further, qRT-PCR results revealed that most of 31 PbrARFs (encoding auxin response factors) in Pyrus bretschneideri were highly expressed in CPF, whereas PbrARF4, PbrARF24 and PbrARF26 were significantly downregulated in CPF vis-a-vis CSF. Phylogenetic analysis revealed that 6 PbrARFs clustered in the group III, where PbrARF4 showed the closest affinity with AtARF1 that promotes organ abscission, indicating a putative role of PbrARF4 in mediating the process of calyx shedding in pear. In fact, the ectopic overexpression of PbrARF4 in Solanum lycopersicum resulted in an earlier-formed and deeper abscission layer (AL) in the transgenic plants, whose calyxes were more prone to wilt at the mature red stage (MR) compared with the control plants (wild-type). More importantly, expression levels of the abscission genes SILS and Sl Cel2 in transgenic plants overexpressing PbrARF4 were significantly upregulated in comparation with the WT, whereas those of Sl BI and Sl TAPG2 were considerably inhibited. Further, PbrJOINTLESS and PbrIDA,the two genes related to calyx shedding in pear, were up-regulated more in CSF than CPF. The findings contribute to a better understanding of PbrARFs involved in fruitlet calyx shedding of pear, which could prove beneficial to improving the quality of pear fruit.

Genomic selection of eight fruit traits in pear

Genomic selection (GS) has the potential to improve selection efficiency and shorten the breeding cycle in fruit tree breeding. In this study,we evaluated the effect of prediction methods, marker density and the training population (TP) size on pear GS for improving its performance and reducing cost. We evaluated GS under two scenarios:(1) five-fold cross-validation in an interspecific pear family;(2) independent validation. Based on the cross-validation scheme, the prediction accuracy (PA) of eight fruit traits varied between 0.33 (fruit core vertical diameter)and 0.65 (stone cell content). Except for single fruit weight, a slightly better prediction accuracy (PA) was observed for the five parametrical methods compared with the two non-parametrical methods. In our TP of 310 individuals, 2 000 single nucleotide polymorphism (SNP) markers were sufficient to make reasonably accurate predictions. PAs for different traits increased by 18.21%-46.98%when the TP size increased from 50to 100, but the increment was smaller (-4.13%-33.91%) when the TP size increased from 200 to 250. For independent validation, the PAs ranged from 0.11 to 0.45 using rrBLUP method. In summary, our results showed that the TP size and SNP numbers had a greater impact on the PA than prediction methods. Furthermore, relatedness among the training and validation sets, and the complexity of traits should be considered when designing a TP to predict the test panel.

Genetic Variation in Resistance to Valsa canker is Related to Arbutin and Gallic Acid Content in Pyrus bretschneideri

Pear Valsa canker is a fungal trunk disease caused by Valsa pyri. Phenolic compounds are ubiquitous in plants and usually contribute to plant resistance against biotic stress. To investigate the association between phenolic compounds and level of resistance to V. pyri, we quantified the contents of individual phenolic compounds in the cortex and phloem of stems from 8 cultivars of Pyrus bretschneideri. Significant variation in the levels of all compounds was found among the cultivars. Correlation analysis revealed an inverse correlation between levels of arbutin and gallic acid with the degree of canker resistance. This suggested that these phenolic compounds are beneficial to V. pyri infection. These data could be valuable for breeding cultivars of P. bretschneideri with greater resistance to V. pyri.

Characterization and Expression Analysis of Pb CS1 Gene Regulated by Iron Deficiency and Involved in Response to Auxin in Pyrus betulifolia

Iron(Fe) is an essential micronutrient required by all fruit trees. To elucidate the regulatory mechanism of auxin to the long-distance transport of Fe, a gene encoding a putative CS protein was isolated from Pyrus betulifolia. This gene was designated as Pb CS1 containing 1 422 bp in length with an open reading frame encoding a protein of 473 amino acids with a predicted molecular mass of 52.69 ku and a theoretical isoelectric point of 6.91. The deduced Pb CS1 protein contained a conserved CS domain and WPNVDAHS sequence existing in the PWPN-box. Based on its conserved domain and phylogenetic status, the Pb CS1 should be grouped into type-Ⅰ subfamily of plant CS. Quantitative real-time PCR results indicated that the Pb CS1 expression was enriched in leaves, roots and phloem, but relatively weak in xylem. Using split root systems, Fe deficiency in one portion of the root system could induce dramatic up-regulation of the Pb CS1 expression in the Fe-sufficient part, suggesting that the Pb CS1 expression was activated by systemic signals. In addition, supplying with NAA and exogenous auxin to the de-topped shoots could recover the Fe deficiency-induced up-regulation of the Pb CS1 expression in the Fe-sufficient part of the root system. In contrast, NPA(an auxin transport inhibitor) application to the shoot tips arrested up-regulation of the Pb CS1 expression in the untreated portion. The results suggested that Fe-deficiency-induced alterations of the Pb CS1 expression were mediated by auxin.

Involvement of long non-coding RNAs in pear fruit senescence under high-and low-temperature conditions

Pear fruit senescence under high-and low-temperature conditions has been reported to be mediated by microRNAs.Long non-coding RNAs(lncRNAs),which can function as competing endogenous RNAs that interact with microRNAs,may also be involved in temperature-affected fruit senescence.Based on the transcriptome and microRNA sequencings,in this study,3330 lncRNAs were isolated from Pyrus pyrifolia fruit.Of these lncRNAs,2060 and 537 were responsive to high-and low-temperature conditions,respectively.Of these differentially expressed lncRNAs,82 and 24 correlated to the mRNAs involved in fruit senescence under high-and low-temperature conditions,respectively.Moreover,three lncRNAs were predicted to be competing endogenous RNAs(ceRNAs)that interact with the microRNAs involved in fruit senescence,while one and two ceRNAs were involved in fruit senescence under high-and low-temperature conditions,respectively.A dual-luciferase assay showed that the interaction of an lncRNA with a microRNA disrupts the action of the microRNA on the expression of its target mRNA(s).Furthermore,four alternative splicing-derived lncRNAs interacted with miR172i homologies(Novel_88 and Novel_69)to relieve the repressed expression of their target and produce an miR172i precursor.Correlation analysis of microRNA expression suggested that Novel_69 is likely involved in the cleavage of the pre-miR172i hairpin to generate mature miR172i.Taken together,lncRNAs are involved in pear fruit senescence under high-or low-temperature conditions through ceRNAs and the production of microRNA.

Comparative genomic analysis of N6-methyladenosine regulators in nine rosaceae species and functional characterization in response to drought stress in pear

N 6-methylated adenine(m6 A)is an emerging epigenetic marker in eukaryotic organisms that plays an important role in biological functions and in enriching genetic information.m6 A exerts these functions via the dynamic interplay among m6 A writers,erasers,and readers.However,little is known about the underlying mechanisms of m6 A in plant growth and stress responses.Here,we identified 276 masked m6 A regulators from nine Rosaceae species(Pyrus bretschneideri,Pyrus betulifolia,Pyrus communis,Malus domestica,Fragaria vesca,Prunus avium,Prunus mume,Prunus persica,and Rubus occidentalis).We classified and named these genes in more detail based on phylogenetic and synteny analysis.The expansion of m6 A regulators in Maloideae was dated back to the recent whole-genome duplication(WGD)in Rosaceae.Based on the expression pattern analysis and gene structure analysis of m6 A regulators,m6 A was shown to be a significant factor in regulating plant development and resistance.In addition,PbrMTA1-silenced pear plants displayed significantly reduced drought tolerance and chlorophyll content,as well as increased electrolyte leakage and concentrations of malondialdehyde and H2 O2.

Mulching broad ridges with a woven polypropylene fabric increases the growth and yield of young pear trees ‘Yuluxiang' in the North China Plain

Unlike traditional ridging, mulching broad ridges with a woven polypropylene fabric (WPF) can reduce soil evaporation during the drought season and avoid long saturation time in the root zone of pear trees during the rainy season. In this study, field experiments were conducted from 2017 to 2020 in a pear orchard in the North China Plain to investigate the effects of mulching broad ridges (0.3 m in height and 2 m in width) with WPF on soil temperature and moisture, nitrogen leaching, vegetative and reproductive growth of young pear trees(Pyrus bretschneideri Rehd.‘Yuluxiang’). The experiments involved two treatments, namely, control (traditional no-ridge planting without mulching) and mulching broad ridges with WPF (RM treatment). The results showed that the RM treatment increased soil moisture and temperature and decreased nitrogen leaching, resulting in vigorous growth of the young pear trees. Moreover, the RM treatment increased the tree trunk cross-sectional area and height of the young pear trees by 37%and 8%in 2020, respectively. The nitrate nitrogen content at the soil layer depth of 0-30 cm was significantly higher in the RM than that in control. Furthermore, the RM treatment significantly increased the fruit yield due to larger tree size. In addition, compared with control, significantly higher fruit soluble solid content of RM treatment was detected in 2020. High precipitation (423 mm) occurred during fruit enlargement stage in 2020, RM treatment decreased the rainfall infiltration in the ridge and the soil moisture in root region, resulting in the improvement of fruit quality, compared with control.Therefore, mulching broad ridges with WPF can be implemented to increase soil moisture during drought season, soil temperature, and nitrate nitrogen content, thereby improving the growth and fruit yield of young pear trees. Additionally, it can reduce soil moisture in the root zone during the rainy season and improve the fruit quality of the trees. Finally, it can reduce nitrate nitrogen leaching, thereby reducing environmental pollution.

Genome-wide identification of the mitogen-activated protein kinase kinases in pear and their functional analysis in response to black spot

The mitogen-activated protein kinase(MAPK)cascade is crucial to plant growth,development,and stress responses.MAPK kinases(MAPKK)play a vital role in linking upstream MAPKK kinases(MAPKKK)with the downstream MAPK.Black spot is one of the most serious fungal diseases of pear which is an important part of the fruit industry in China.The MAPKK genes have been identified in many plants,however,none has been reported in pear(Pyrus bretschneideri).In order to explore whether MAPK gene of pear is related to black spot disease,we designed this experiment.The present study investigated eight putative PbrMAPKK genes obtained from the Chinese white pear genome.The phylogenetic analysis revealed that PbrMAPKK genes were divided into A,B,C,and D groups.These PbrMAPKK genes are randomly distributed on 7 out of 17 chromosomes and mainly originated from the whole-genome duplication(WGD)event.The expression analysis of PbrMAPKK genes in seven pear tissues and the leaves of susceptible and resistant varieties after Alternaria alternata infection by quantitative real-time PCR(qRT-PCR)identified seven candidate genes associated with resistance.Furthermore,virus-induced gene silencing(VIGS)indicated that PbrMAPKK6 gene enhanced resistance to pear black spot disease in pear.