Analysis of Subcellular Localization and Pathogenicity of Plum Bark Necrosis Stem-Pitting Associated Virus Protein P6

Infection of plum bark necrosis stem pitting associated virus(PBNSPaV)has been reported in many Prunus species in several countries,causing significant economic losses.The very small proteins encoded by plant viruses are often overlooked due to their short sequences and uncertain significance.However,numerous studies have indicated that they might play important roles in the pathogenesis of virus infection.The role of small hydrophobic protein P6,encoded by the open reading frame 2 of PBNSPaV,has not been well explored.In this study,we amplified the P6 fragment from a PBNSPaV isolate by RT-PCR using specific primers and found that it is 174 bp long and encodes a protein of approximately 6.3 kD with a transmembrane domain.Subcellular localization analysis of P6 proteins in tobacco leaves showed that P6 localizes to the cytomembrane and nuclear membrane.To further clarify the pathogenicity of P6 proteins,we constructed a PVX-P6 expression vector by inserting the p6 fragment into a potato virus X(PVX)-based vector and transformed it into Agrobacterium tumefaciens GV3101.Infiltration of Nicotiana benthamiana(N.benthamiana)with the PVX vector-transformed A.tumefaciens led to slight mosaic symptoms at 14 days of post-inoculation.Meanwhile,infiltration with the PVX-P6 vector-transformed A.tumefaciens resulted in no significant symptoms.These results demonstrated that heterologous expression of P6 in N.benthamiana could not enhance the pathogenicity of PVX.Our study indicates that P6 may not be a potential pathogenic factor associate with the causing of symptoms,and the mode of action of PBNSPaV-P6 protein remains to be further studied.

经腹入路3D腹腔镜联合输尿管硬镜同期处理肾盂输尿管连接部梗阻并肾盏结石的应用

目的探讨经腹入路3D腹腔镜联合输尿管硬镜在肾盂输尿管连接部梗阻并肾盏结石手术中的安全性及有效性。方法回顾性分析2019年12月至2022年1月广东省第二人民医院收治并手术的14例肾盂输尿管连接部梗阻并肾盏结石的患者临床资料,分析肾盏结石大小、结石数量、手术时间、术中失血量及一期结石清除率及手术效果。结果所有患者均采用经腹3D腹腔镜下肾盂离断成形术,术中输尿管镜经腔镜通道进入肾盂肾盏找到结石,取出结石。结石大小(0.89±0.32) cm,结石数量2~6枚,手术时间(138±18)min,术中失血量(12±5)ml,一期结石清除率100%,术后6~8周拔除输尿管支架管,术后3个月复查CT示肾积液缓解或消失,无肾结石复发及肾积液加重。结论3D腹腔镜联合输尿管硬镜在肾盂输尿管连接部梗阻并肾盏结石治疗中安全有效,具有创伤小,清石率高,是部分肾盂输尿管连接部梗阻合并肾盏结石患者治疗的良好选择。

Altitude-dependent metabolite biomarkers reveal the mechanism of plateau pika adaptation to high altitudes

The harsh environment in the Tibetan plateau,the highest place in the world,poses thermoregulatory challenges and hypoxic stress to animals.The impacts of plateau environment on animal physiology and reproduction include external factors such as strong ultraviolet radiation and low temperature,and internal factors such as animal metabolites and gut microbiota.However,it remains unclear how plateau pika adapt to high altitudes through the combination of serum metabolites and gut microbiota.To this end,we captured 24 wild plateau pikas at the altitudes of 3400,3600,or 3800 m a.s.l.in a Tibetan alpine grassland.Using the machine learning algorithms(random forest),we identified five biomarkers of serum metabolites indicative of the altitudes,that is,dihydrotestosterone,homo-l-arginine,alpha-ketoglutaric-acid,serotonin,and threonine,which were related to body weight,reproduction,and energy metabolism of pika.Those metabolic biomarkers were positively correlated with Lachnospiraceae_Agathobacter,Ruminococcaceae,or Prevotellaceae_Prevotella,suggesting the close relationship between metabolites and gut microbiota.By identifying the metabolic biomarkers and gut microbiota analysis,we reveal the mechanisms of adaptation to high altitudes in plateau pika.

A positive feedback circuit for ROP-mediated polar growth

Tip growth is a special type of polarized growth in which a single and unique polarization site is established and maintained.Rho of Plants(ROP)proteins,which represent the only class of Rho GTPases in plants,regulate tip growth.The dynamic and asymmetric distribution of ROPs is critical for the establishment and maintenance of tip growth,and requires at least one positive feedback loop,which is still elusive.Here,we report a positive feedback circuit essential for tip growth of root hairs,in which ROPs,ROP activators and effectors,and AGC1.5 subfamily kinases are interconnected by sequential oligomerization and phosphorylation.AGC1.5 subfamily kinases interact with and phosphorylate two guanine nucleotide exchange factors(GEFs)of ROPs,RopGEF4 and RopGEF10.They also interact with two ROP effectors,ICR2/RIP3 and MIDD1/RIP4,which bridge active ROPs with AGC1.5.Functional loss of the AGC1.5 subfamily kinases or ICR2 and MIDD1 compromised root hair growth due to reduced ROP signaling.We found that asymmetric targeting of RopGEF4 and RopGEF10 is controlled by AGC1.5-dependent phosphorylation.Interestingly,we discovered that the ROP effectors recruit AGC1.5 to active ROP domains at the plasma membrane during root hair growth and are critical for AGC1.5-dependent phosphorylation of RopGEFs.Given the large number of AGC kinases in plants,this positive feedback circuit may be a universal theme for plant cell polar growth.

Influence analysis of sprinkler irrigation effectiveness using ANFIS

In order to improve sprinkler irrigation quality and promote actual irrigation efficiency,the influence analysis of sprinkler irrigation effectiveness(SIE)using ANFIS(Adaptive Neural Fuzzy Inference System)was implemented to balance moisture infiltration and water redistribution in soil field.Firstly,using a detailed description of governing equations proposed for sprinkler irrigation flow,the theoretical foundation and mathematical model of irrigation effectiveness can be established;Secondly,based on a complete preparation of experimental irrigation conditions,a series of calibration indexes quantifying SIE for sprinkler irrigation quality and infiltration efficiency were proposed;Then thirdly,a novel ANFIS system was designed and introduced to evaluate these key effectiveness indexes in actual working operations,so that a series of detailed influence analysis and comprehensive infiltration assessment focusing on sprinkler irrigation effectiveness could be achieved afterwards,which result to the realization of better infiltration equilibrium and higher water redistribution efficiency in actual irrigation test.Therefore,the qualification of sprinkler irrigation effectiveness was achieved,and in addition,the moisture infiltration improvement and soil moisture uniformity were facilitated also in return.