Informing native plant sourcing for ecological restoration: cold‐hardiness dynamics,flowering phenology,and survival of Eriogonum umbellatum

Despite advances in restoration of degraded lands around the world, native plants are still underutilized. Selection of appropriate plant materials is a critical factor in determining plant establishment and persistence. To better inform decision‐making, we examined cold‐hardiness dynamics, flowering phenology, and survival among five geographically distinct sulfur‐flower buckwheat (Polygonaceae: Eriogonum umbellatum Torr.) populations in a common garden. LT₅₀ (a measure of freezing injury) was determined every 6 weeks across a complete year; one population was also evaluated at the source. Cold‐hardiness dynamics were similar across populations, with annual fluctuations in mean LT₅₀ exceeding 40°C. Rate of deacclimation (i.e. loss of cold tolerance) in spring varied across populations and was not related to the elevation from which a population came. Plants were less cold hardy in October 2014 compared to October 2013, likely reflecting a response to colder local conditions in 2013. Although the range of LT₅₀ was similar for a single comparison of common garden versus wild‐grown plants, wild‐grown plants acclimated and deacclimated earlier than common garden‐grown plants. Plants derived from a low‐elevation population showed delayed flowering phenology, while high‐elevation populations showed earlier flowering phenology, with one high‐elevation population having the lowest survival rate in the common garden. These results suggest that while considerable plasticity in seasonal cold‐hardiness dynamics occur, population variability in deacclimation and flowering phenology have implications for selection and movement of sulfur‐flower buckwheat for ecological restoration.     

叶面喷施PP_(333)和6-BA对甜荞花芽分化及内源激素的影响

以甜荞品种‘北早生’和‘赤甜荞1号’为材料,利用多效唑(PP_(333))和6-苄基腺嘌呤(6-BA)在不同幼苗时期进行叶片喷施处理,并采用扫描电镜观察花芽分化过程,记录开花时间,调查开花和结实数,通过液质联用方法检测3种内源激素生长素(IAA)、赤霉素(GA_3)、脱落酸(ABA)及人工合成细胞分裂素(6-BA)含量,探究两种植物生长调节剂对甜荞的花芽分化及内源激素含量的影响,为甜荞花期调控提供依据。结果显示:(1)第二片真叶期喷施100mg·L~(-1)PP_(333)和150mg·L~(-1)6-BA能显著提高甜荞结实率和单株产量,两种处理的结实率分别较对照提高49.5%、39.4%,单株产量分别提高41.8%、23.0%。(2)观察甜荞花芽分化过程并将其划分5个时期,分别为生长锥分化前期、生长锥分化期、小花原基分化期、雌雄蕊分化期和雌雄蕊形成期,在第二片真叶期喷施PP_(333)和6-BA均加快了甜荞花芽分化进程,使其开花时间提前。(3)喷施PP_(333)在幼苗期和现蕾期降低了内源IAA、6-BA含量,增加GA_3、ABA含量,而在盛花期增加内源IAA、6-BA含量,降低ABA、GA_3含量;喷施6-BA在幼苗期和现蕾期增加了内源GA_3、IAA含量,在盛花期降低了内源GA_3、IAA含量,3个时期均降低了内源ABA含量,增加了6-BA含量。研究发现,第二片真叶期是调控甜荞小花发育的关键时期,在此时叶面喷施100mg·L~(-1)PP_(333)和150mg·L~(-1)6-BA的调控效果最佳,可加快甜荞花芽分化进程,使开花时间提前,增加单株结实粒数,提高甜荞品种的结实率和单株产量;PP_(333)主要是通过减少甜荞开花数,增强弱势小花活力来提高结实粒数,而6-BA主要是利用增加开花数,提高有效可孕小花数来增加结实粒数。     

苦荞R2R3-MYB转录因子调控原花青素生物合成的研究

基于苦荞花期转录组数据,该研究筛选并克隆获得一个黄酮代谢相关的MYB类转录因子,并命名为FtMYB23。该基因ORF框长879bp,编码292个氨基酸;系统进化树分析显示,FtMYB23与SG5-MYB亚家族成员聚为一簇,属于典型的R2R3-MYB型转录因子。β-半乳糖苷酶滤纸分析表明,其具有转录激活活性。FtMYB23过表达转基因拟南芥株系的表型分析表明,3个阳性株系的种皮颜色均呈现出比野生型更深的褐色,其叶中原花青素含量均极显著增加(P 0.01),分别为野生型的4.68、3.5和2.8倍。qRT-PCR分析表明,转基因拟南芥中黄酮合成相关的AtCHS、AtCHI、AtF3H、AtF3′H、AtFLS、AtDFR和AtBAN等基因的表达量显著升高(P 0.05),而AtTT12的表达量极显著降低(P 0.01)。研究认为,FtMYB23作为典型的Subgroup5-MYB(SG5-MYB)激活型转录因子,通过促进黄酮合成途径早期关键酶基因的表达,从而提高原花青素的合成与积累。     

The effector SIX8 is required for virulence of Fusarium oxysporum f.sp. cubense tropical race 4 to Cavendish banana

Plant pathogens employ effectors as molecular weapons to manipulate host immunity and facilitate colonization. Fusarium oxysporum f. sp. cubense is the agent of wilt disease in banana plantlets and four races of the pathogen have been identified based on the cultivar specificity. A total of 9 SIX genes have been detected in the genome of Foc TR4 and 6 genes detected in Foc1. Among these SIX genes, SIX2 and SIX8 are only detected in Foc TR4, not identified in Foc1. Expression profiles analysis revealed that SIX genes of Foc TR4 are highly induced after inoculation to Cavendish banana plantlets. Virulence analysis of the SIX2 and SIX8 knock-out mutants showed that SIX8 is required for the virulence of Foc TR4 while SIX2 has no obvious functions. Over expression of SIX8-FLAG proteins in the SIX8 knock-out mutant partly restored the virulence. Western blot analysis suggested that SIX8 could be secreted into the extracellular space and a signal peptide resided the N-terminal polypeptide sequence. This study provides some clues for further research on mechanism of SIX8 in regulating virulence of Foc TR4.     

Agrilus mali Matsumara (Coleoptera: Buprestidae), a new invasive pest of wild apple in western China: DNA barcoding and life cycle

Agrilus mali Matsumara (Coleoptera: Buprestidae) is a wood‐boring beetle distributed to eastern China that occasionally injures apple species. However, this wood‐boring beetle is new to the wild apple forests (Malus sieversii) of the Tianshan Mountains (western China) and has caused extensive tree mortality. The development of a biological control program for these wild apple forests is a high priority that requires exploration of the life cycle, DNA barcoding and taxonomic status of A. mali. In this study, to determine the diversity of invasive beetles, a fragment of the mitochondrial cytochrome oxidase gene was analyzed. Based on the results, beetles from Gongliu and Xinyuan counties of Xinjiang were identical but differed from those in the apple nursery of Gongliu by a single‐nucleotide substitution. We summarize the taxonomic status, relationships, and genetic distances of A. mali among other Agrilus species using the Tajima‐Nei model in maximum likelihood phylogeny. Analysis revealed that A. mali was closely related to Agrilus mendax and both belong to the Sinuatiagrulus subgenus. The life cycle of A. mali was investigated based on a monthly regular inspection in the wild apple forests of Tianshan. Similar to congeneric species, hosts are injured by larvae of A. mali feeding on phloem tissue, resulting in serpentine galleries constructed between bark and xylem that prevent nutrient transport and leading to tree mortality. Future studies will focus on plant physiological responses to the invasive beetles and include surveys of natural enemies for a potential classical biological control program.     

Potential benefits and harms: a review of poisonous mushrooms in the world

Mushrooms have long been considered as delicacies as well as used as important dietary supplements and food. However, there are major concerns with poisonous mushrooms as these pose threats to public health and safety. In this paper, we provide a review focusing on poisonous mushrooms, their toxins, symptoms and utilizations. In addition, this paper establishes a poisonous mushroom list which includes 643 species from two phyla, 16 orders, 51 families and 148 genera. The toxicity of all these species was verified and 332 species were ranked as P1 signifying that these species have toxic studies and or clinical poisoning case records and 311 species were P2 meaning they had previously been recorded as poisonous in other studies. Furthermore, we discuss advances in technology including how genomic studies could be used as a breakthrough tool in the field of toxic mushrooms. With this comprehensive review, we aim to promote public awareness of poisonous mushrooms, including how to avoid mushroom poisoning, and how to better utilize poisonous mushroom resources.     

The Cbl-interacting protein TULA inhibits dynamin-dependent endocytosis

The Cbl- and ubiquitin-interacting protein T-cell ubiquitin ligand (TULA) has been demonstrated to inhibit endocytosis and downregulation of ligand-activated EGF receptor (EGFR) by impairing Cbl-induced ubiquitination. We presently report that TULA additionally inhibited clathrin-dependent endocytosis in general, as both uptake of transferrin (Tf) and low-density lipoprotein (LDL) was inhibited. Additionally, endocytosis of the raft proteins CD59 and major histocompatibility complex class I (MHC-I), which we demonstrate were mainly endocytosed clathrin-independently, but dynamin-dependently, was blocked in cells overexpressing TULA. By contrast, the uptake of ricin, which is mainly endocytosed clathrin- and dynamin-independently, was not affected by overexpressed TULA. Consistently, TULA and dynamin co-immunoprecipitated and colocalized intracellularly, and upon overexpression of dynamin the TULA-mediated inhibitory effect on endocytosis of Tf, LDL, CD59 and MHC-I was counteracted. Overexpressed dynamin did not restore ubiquitination of the EGFR, and consistently dynamin did not rescue endocytosis of the EGFR in cells overexpressing TULA. We conclude that TULA inhibits both clathrin-dependent and clathrin-independent endocytic pathways by functionally sequestering dynamin via the SH3 domain of TULA binding proline-rich sequences in dynamin.     

Single amino acid substitution in HIV-1 integrase catalytic core causes a dramatic shift in inhibitor selectivity

HIV-1 integrase (IN) mediates the insertion of viral cDNA into the cell genome, a vital process for replication. This step is catalyzed by two separate DNA reaction events, termed 3'-processing and strand transfer. Here, we show that six inhibitors from five structurally different classes of compounds display a selectivity shift towards preferential strand transfer inhibition over the 3'-processing activity of IN when a single serine is substituted at position C130. Even though IN utilizes the same active site for both reactions, this finding suggests a distinct conformational dissimilarity in the mechanistic details of each IN catalytic event.     

Development of Oryza rufipogon and O. sativa Introgression Lines and Assessment for Yield-related Quantitative Trait Loci

Introgression lines population was effectively used in mapping quantitative trait loci （QTLs）, identifying favorable genes, discovering hidden genetic variation, evaluating the action or interaction of QTLs in multiple conditions and providing the favorable experimental materials for plant breeding and genetic research. In this study, an advanced backcross and consecutive selfing strategy was used to develop introgression lines （ILs）, which derived from an accession of Oryza rufipogon Griff. collected from Yuanjiang County, Yunnan Province of China, as the donor, and an elite indica cultivar Teqing （O. sativa L.）, as the recipient. Introgression segments from O. rufipogon were screened using 179 polymorphic simple sequence repeats （SSR） markers in the genome of each IL. Introgressed segments carried by the introgression lines population contained 120 ILs covering the whole O. rufipogon genome. The mean number of homozygous O. rufipogon segments per introgression line was about 3.88. The average length of introgressed segments was approximate 25.5 cM, and about 20.8% of these segments had sizes less than 10 cM. The genome of each IL harbored the chromosomal fragments of O. rufipogon ranging from 0.54% to 23.7%, with an overall average of 5.79%. At each locus, the ratio of substitution of O. rufipogon alleles had a range of 1.67-9.33, with an average of 5.50. A wide range of alterations in morphological and yield-related traits were also found in the introgression lines population. Using single-point analysis, a total of 37 putative QTLs for yield and yield components were detected at two sites with 7%-20% explaining the phenotypic variance. Nineteen QTLs （51.4%） were detected at both sites, and the alleles from O. rufipogon at fifteen loci （40.5%） improved the yield and yield components in the Teqing background. These O. rufipogon-O, sativa introgression lines will serve as genetic materials for identifying and using favorable genes from common wild rice.     

Expression of a buckwheat trypsin inhibitor gene in Escherichia coli and its effect on multiple myeloma IM-9 cell proliferation

The gene of buckwheat trypsin inhibitor （BTI） has been cloned and expressed in Escherichia coli. The yield of this recombinant inhibitor was over 12 mg/L by using one-step purification on a Ni^2＋-NTA Sepharose column. Its molecular weight was 9322.1 Da, determined by mass spectrum analysis. The MTT and cytometry analyses showed that recombinant BTI could specifically inhibit the proliferation of IM-9 human B lymphoblastoid ceils （from patient with multiple myeloma） in a dose-dependent manner. The test of recombinant BTI-induced apoptosis in IM-9 ceils implied that the inhibitor might have potential application in the treatment of cancer.