Using an ecophysiological analysis to dissect genetic variability and to propose an ideotype for nitrogen nutrition in pea

BACKGROUNDS AND AIMS: Nitrogen nutrition of legumes, which relies both on atmospheric N2 and soil mineral N, remains a major limiting factor of growth. A decade ago, breeders tried to increase N uptake through hypernodulation. Despite their high nodule biomass, hypernodulating mutants were never shown to accumulate more nitrogen than wild types; they even generally displayed depressed shoot growth. The aim of this study was to dissect genetic variability associated with N nutrition in relation to C nutrition, using an ecophysiological framework and to propose an ideotype for N nutrition in pea. METHODS: Five pea genotypes (Pisum sativum) characterized by contrasting root and nodule biomasses were grown in the field. Variability among genotypes in dry matter and N accumulation was analysed, considering both the structures involved in N acquisition in terms of root and nodule biomass and their efficiency, in terms of N accumulated through mineral N absorption or symbiotic N2 fixation per amount of root or nodule biomass, respectively. KEY RESULTS: Nodule efficiency of hypernodulating mutants was negatively correlated to nodule biomass, presumably due to the high carbon costs induced by their excessive nodule formation. Root efficiency was only negatively correlated to root biomass before the beginning of the seed-filling stage, suggesting competition for carbon between root formation and functioning during the early stages of growth. This was no longer the case after the beginning of the seed-filling stage and nitrate absorption was then positively correlated to root biomass. CONCLUSIONS: Due to the high C costs induced by nodule formation and its detrimental effect on shoot and root growth, selecting traits for the improvement of N acquisition by legumes must be engineered (a) considering inter-relationships between C and N metabolisms and (b) in terms of temporal complementarities between N2 fixation and nitrate absorption rather than through direct increase of nodule and/or root biomass.     

Micropropagation and germplasm conservation of Central Park Splendor Chinese elm (<Emphasis Type="Italic">Ulmus parvifolia</Emphasis> Jacq. ‘A/Ross Central Park’) trees

Micropropagation offers opportunities to propagate, preserve and ship tree germplasm. It also reduces the risk of moving pathogens and insects with the germplasm due to built-in pathogen detection capabilities of aseptic cultures. For the past few decades, our laboratory has been involved in a project to preserve and restore a large, cold hardy, and historically important Chinese elm (Ulmus parvifolia Jacq. ‘A/Ross Central Park’) tree. Here we present three simple and efficient systems for its micropropagation, germplasm conservation and distribution: (1) in vitro plant formation from meristematic nodules (MNs), (2) plantlet generation from axillary buds, and (3) in vitro rooting of micro-cuttings from 20-years-old hedged stock plants. Newly flushed nodal segments were used as explants. WPM with 0.5 mg/l BA was found to be the best medium for meristematic shoot development and WPM supplemented with 2.0 mg/l 4-CPPU and 0.5 mg/l TDZ was best for meristematic nodule formation. Rhizogenesis of regenerants and micro-cuttings was best achieved on WPM with 1.0 mg/l NAA and 2% sucrose. Rooted plants were readily acclimatized to the greenhouse ambient environment and continued to grow well under greenhouse conditions. The survival rate of acclimatized plantlets under ex vitro conditions was 100% after 4 weeks. Plants looked healthy with no visually detectable phenotypic variation based on observation of about 1,000 plants. Cycling of shoot explants and MNs through repetitive cultures was effective in scaling-up propagules.     

声触诊组织定量成像联合声诺维超声造影对乳腺BI-RADS 4类结节的鉴别诊断价值

摘要 目的：探讨声触诊组织定量成像（VTIQ）联合声诺维超声造影对乳腺BI-RADS 4类结节的鉴别诊断价值。方法：将我院2018年6月至2019年12月门诊及住院诊治的90例女性乳腺结节患者作为研究对象,利用VTIQ超声弹性成像技术,获取剪切波速度最大值（SWVmax）、剪切波速度最小值（SWVmin）、剪切波速度平均值（SWVmean）,声诺维超声造影评估病灶回声、形态、血管充盈情况。以手术病理或超声引导穿刺活检病理结果作为金标准,分析VTIQ超声弹性成像联合声诺维超声造影对乳腺BI-RADS 4类结节的鉴别诊断价值。结果：90例患者（90个乳腺结节）中,经手术病理检查62例,经超声引导穿刺活检28例,其中良性结节48例,恶性结节42例。乳腺BI-RADS 4类良性及恶性结节的SWVmax、SWVmin及SWVmean比较有统计学差异（P<0.05）;以SWVmax≥3.83 m/s作为截断点,鉴别诊断乳腺BI-RADS 4类良恶性结节的曲线下面积（AUC）、敏感度、特异度分别为0.875、83.33%、91.67%,均高于SWVmin及SWVmean。SWVmax、声诺维超声造影与联合诊断的准确率比较无统计学差异（P>0.05）。SWVmax联合声诺维超声造影检查在鉴别诊断乳腺BI-RADS 4类良恶性结节的AUC、敏感度、特异度分别为0.914、95.24%、87.50%,诊断效能最佳。结论：声诺维超声造影、VTIQ超声弹性成像在乳腺BI-RADS 4类结节鉴别诊断中有一定的价值,VTIQ超声弹性成像的SWVmax参数诊断效能优于SWVmin及SWVmean,VTIQ超声弹性成像联合声诺维超声造影可提升乳腺BI-RADS 4类结节鉴别诊断效能。     

施肥对红豆树苗木生长、根系结瘤和矿质营养的影响

探讨几种市售肥料对红豆树(Ormosia hosiei)苗木各器官生长发育及养分含量的影响,为培育红豆树优质苗木选用肥料提供参考。以红豆树当年生大田苗为试材,采用3种常见肥料(复合肥、缓释肥和尿素)分3个施肥水平进行施肥试验,测定苗高、地径、根系指标、根系结瘤指标、各器官生物量及N、P、K含量,对不同施肥处理苗木质量进行综合评价。不同施肥处理对红豆树苗木生长、结瘤和矿质营养均有显著或极显著影响。施缓释肥和复合肥的处理表现为随施肥量的增加苗木的生长和生理指标均表现出先增后降趋势,施尿素则表现为随施肥量增加呈逐渐递减趋势。通过隶属函数对苗木质量进行综合评价,表明缓释肥30 g·m^-2为最佳施肥处理,其苗高增量、各根系质量指标、各生物量指标、结瘤数量以及根、茎、叶的N、P、K含量均为最大。红豆树当年生苗木密度为50 株·m-2时,施用缓释肥(N-P-K: 30-6-7) 30 g·m^-2可以有效提高红豆树的苗木质量。     

Gibberellin controls the nodulation signaling pathway in Lotus japonicus

Root nodule formation is regulated by several plant hormones, but the details of the regulation of the nodulation signaling pathway are largely unknown. In this study, the role of gibberellin (GA) in the control of root nodule symbiosis was investigated at the physiological and genetic levels in Lotus japonicus . Exogenous application of biologically active GA, GA₃, inhibited the formation of infection threads and nodules, which was counteracted by the application of a biosynthesis inhibitor of GA, Uniconazole P. Nod factor-induced root hair deformation was severely blocked in the presence of GA, which was phenocopied by nsp2 mutants. The number of spontaneous nodules triggered by the gain-of-function mutation of calcium/calmodulin-dependent kinase (CCaMK) or the lotus histidine kinase 1 (LHK1) was decreased upon the addition of GA; moreover, the overexpression of the gain-of-function mutation of L. japonicus , SLEEPY1, a positive regulator of GA signaling, resulted in a reduced nodule number, without other aspects of root development being affected. These results indicate that higher GA signaling levels specifically inhibit the nodulation signaling pathway. Nod factor-dependent induction of NSP2 and NIN was inhibited by exogenous GA. Furthermore, the cytokinin-dependent induction of NIN was suppressed by GA. From these results, we conclude that GA inhibits the nodulation signaling pathway downstream of cytokinin, possibly at NSP2, which is required for Nod factor-dependent NIN expression. These results clarify the roles of GA in the nodulation signaling pathway, and in relation to the cytokinin signaling pathway for nodulation in L. japonicus .     

Diversity of frankiae in soils from five continents

Clone libraries of nifH gene fragments specific for the nitrogen-fixing actinomycete Frankia were generated from six soils obtained from five continents using a nested PCR. Comparative sequence analyses of all libraries (n=247 clones) using 96 to 97% similarity thresholds revealed the presence of three and four clusters of frankiae representing the Elaeagnus and the Alnus host infection groups, respectively. Diversity of frankiae was represented by fewer clusters (i.e., up to four in total) within individual libraries, with one cluster generally harboring the vast majority of sequences. Meta-analysis including sequences previously published for cultures (n=48) and for uncultured frankiae in root nodules of Morella pensylvanica formed in bioassays with the respective soils (n=121) revealed a higher overall diversity with four and six clusters of frankiae representing the Elaeagnus and the Alnus host infection groups, respectively, and displayed large differences in cluster assignments between sequences retrieved from clone libraries and those obtained from nodules, with assignments to the same cluster only rarely encountered for individual soils. These results demonstrate large differences between detectable Frankia populations in soil and those in root nodules indicating the inadequacy of bioassays for the analysis of frankiae in soil and the role of plants in the selection of frankiae from soil for root nodule formation.     

Endophytic occupation of peanut root nodules by opportunistic Gammaproteobacteria

Several bacterial isolates were recovered from surface-sterilized root nodules of Arachis hypogaea L. (peanut) plants growing in soils from Córdoba, Argentina. The 16S rDNA sequences of seven fast-growing strains were obtained and the phylogenetic analysis showed that these isolates belonged to the Phylum Proteobacteria, Class Gammaproteobacteria, and included Pseudomonas spp., Enterobacter spp., and Klebsiella spp. After storage, these strains became unable to induce nodule formation in Arachis hypogaea L. plants, but they enhanced plant yield. When the isolates were co-inoculated with an infective Bradyrhizobium strain, they were even found colonizing pre-formed nodules. Analysis of symbiotic genes showed that the nifH gene was only detected for the Klebsiella-like isolates and the nodC gene could not be amplified by PCR or be detected by Southern blotting in any of the isolates. The results obtained support the idea that these isolates are opportunistic bacteria able to colonize nodules induced by rhizobia.     

Organization of the lamin scaffold in the internal nuclear matrix of normal and transformed hepatocytes

Nuclear lamins are among the more abundant proteins making up the internal nuclear matrix, but very little is known about their structure in the nucleoplasm. Using immunoelectron microscopy, we demonstrate the organization of lamins in the nuclear matrix isolated from rat hepatocytes for the first time. Lamin epitopes are arrayed both in locally ordered clusters and in quasi-regular rows. Fourier filtering of the images demonstrates that the epitopes are placed at the nodes and halfway between the nodes of square or rhombic lattices that are about 50 nm on each side, as well as along rows at regular ∼25-nm intervals. In addition, we have compared this structure with that of the internal nuclear matrix isolated from persistent hepatocyte nodules. In transformed hepatocytes, the islands of lamin lattice are lost, and only a partial regularity in the rows of gold particles remains. We suggest that orthogonal lattice assembly might be an intrinsic property of lamin molecules, and that the disassembly may be triggered by simple molecular events such as phosphorylation.     

虚拟导航支气管镜联合EBUS-GS技术在直径≤2 cm的肺外周结节诊断中的应用

摘要 目的：探究虚拟导航支气管镜联合EBUS-GS技术在直径≤2 cm的肺外周结节诊断中的诊断价值。方法：本研究采用回顾性分析的方法,选取2019年5月-2022年5月期间在我院就诊的140例直径≤2 cm的肺外周结节患者作为研究对象,随机分为对照组和实验组,各70例。对照组使用传统支气管镜技术进行诊断,实验组使用虚拟导航支气管镜联合EBUS-GS技术进行诊断,并进行随访实验。记录并比较两组患者的可视率、操作时间、诊断率及并发症发生率等指标。结果：对照组和实验组中的病灶位置比较,两组差异无统计学意义（P>0.05）。实验组与对照组病变大小相比差异无统计学意义（P>0.05）,实验组可视率高于对照组。对照组和实验组的支气管镜操作总时间比较,无明显统计学差异（P>0.05）,探及病灶位置时间与对照组相比,实验组低于对照组（P<0.05）。实验组中恶性病变的诊断率为89.74%,良性肺结节的诊断率为83.87%,而对照组恶性病变的诊断率为60%,良性结节的诊断率为50%,两组间病变性质的良性和恶性诊断率相比较,差异均具有统计学意义（P<0.05）。与对照组相比,实验组不良反应发生率2.86%低于对照组不良反应发生率18.57%,差异具有统计学意义（P<0.05）。结论：虚拟导航支气管镜联合EBUS-GS技术是一种可行且有效的方法,可用于直径≤2 cm的肺外周结节的诊断。该技术既可提高诊断准确性,又可减少不必要的切除手术,对患者具有重要的临床意义。     

Effect of water stress on the enzymes of nitrogen metabolism in mung bean (Vigna radiata Wilczeck) nodules

Abstract. Water stress created by withholding irrigation in mung bean resulted in decreased leaf water potential and nodule moisture content. Decreased leaf water potential was associated with decreased activity of nitrogenase, glutamine synthetase (GS), asparagine synthetase (AS), aspartate amino transferase (AAT), xanthine dehydrogenase (XDH) and uricase. However, the activity of glutamate dehydrogenase increased three-fold under severe stress. The activity of allantoinase and allantoicase was not affected by moderate stress but decreased under severe stress. The in vitro production of allantoic acid from allantoin and uric acid in the cytosol fraction decreased more than its production from xanthine and hypoxanthine. The production of NADH also decreased under stress.
During recovery from severe stress, the activity of XDH and uricase further decreased, whilst that of allantoinase and allantoicase increased compared to the control. This corresponded with the higher content of ureides during recovery. The recovery in other enzymes was not complete although leaf water potential and nodule moisture content recovered fully within 24 h.