磁共振成像技术在毛竹幼竹显微结构和水分含量表征中的应用

竹子节部对竹秆的中空结构以及竹秆快速高生长起着关键作用,研究竹子解剖结构有助于认识竹子生长机制。本研究利用核磁共振成像(MRI)无创、高分辨率和准确的技术优势,对毛竹幼竹去笋箨梢部进行横切面高精度核磁共振成像扫描,使用MATLAB软件对 MRI进行灰度值采集,分析节部、近节部和节间水分分布差异。结果表明: 无数维管束在毛竹幼竹节隔以及近节隔内腔反复扭曲和水平转动,组成了一个错综复杂和高度连接的网络结构,通过分轴向载荷来保护重要组织免受机械应力,同时实现水分和营养物质的横向运输,这是毛竹在短时间内快速完成高生长的重要基础。MRI信号值(亮度值)表明,幼竹维管束的水分含量远远高于周围的基本组织。节间平均水分含量和像素点间含水量标准差显著高于节部,近节部居中。MRI技术可以在未来竹子解剖学和生理生化学研究中发挥作用。     

毛竹GRF转录因子家族的全基因组鉴定与分析

利用生物信息学方法,于毛竹(Phyllostachys edulis (Carr.) Lehaie)全基因组中鉴定获得18个GRF转录因子,并对其理化特性、保守结构域、系统发育关系、mi R396靶位点以及基因表达模式进行了分析。结果表明,18个Pe GRF蛋白长度为170～551 aa,分子量为18.5～58.8 k D;这些Pe GRF蛋白均具有QLQ和WRC结构域,部分Pe GRF含有FFD和TQL保守结构域。对毛竹、拟南芥(Arabidopsis thaliana (L.) Heynh)和水稻(Oryza sativa L.)的系统进化分析结果显示,毛竹18个GRF可分为3个亚类,且单子叶植物毛竹和水稻的GRF转录因子亲缘关系更近。mi R396靶位点预测分析结果发现,在13个Pe GRF基因序列的编码区存在毛竹mi R396结合位点; Pe GRF基因表达模式分析结果显示,Pe GRF主要在毛竹的竹笋中表达。     

Analysis of the expression of <Emphasis Type="Italic">BohLOL1</Emphasis>, which encodes an LSD1-like zinc finger protein in <Emphasis Type="Italic">Bambusa oldhamii</Emphasis>

A cDNA, BohLOL1, encoding a protein containing three zf-LSD1 (zinc finger-Lesions Simulating Disease resistance 1) domains was cloned from growing bamboo (Bambusa oldhamii) shoots. A phylogenetic analysis revealed that BohLOL1 is a homolog of Arabidopsis LSD1 and LOL1 (LSD-one-like 1), which have been reported to act antagonistically in controlling cell death via the maintenance of reactive oxygen species homeostasis. The BohLOL1 gene was differentially expressed in various bamboo shoot tissues and was upregulated in shoots with higher rates of culm elongation. The expression level of this gene in multiple shoots of bamboo, which were cultured in vitro, was also upregulated by auxins, cytokinins, pathogen infection, 2,6-dichloroisonicotinic acid (a functional analog of salicylic acid), and hydrogen peroxide. The results suggest that BohLOL1 participates in bamboo growth and in the response to biotic stress. The DNA-binding assays and subcellular localization studies demonstrated that BohLOL1 is a nuclear DNA-binding protein. BohLOL1 might function through protein-DNA interactions and thus affect the expression of its target genes. The results of this study extend the role of plant LSD1 and LOL1 proteins from the regulation of cell death to cell growth. The growth-dependent up-regulation of BohLOL1 expression, which uniquely occurs in growing bamboo, might be one of the critical factors that contribute to the rapid growth of this remarkable plant.     

四个竹秆变异毛竹变型的全基因组序列分析

毛竹是我国重要的经济竹种,在长期栽培适应过程中产生了丰富的变异。为揭示毛竹竹秆变异变型的全基因组突变类型,以黄皮毛竹、金丝毛竹、绿皮花毛竹和花毛竹4个毛竹变型为实验材料,采用高通量重测序技术获得全基因组序列,进行单核苷多态性(SNP)、小片段插入缺失(InDel)和结构变异(SV)检测和注释,并将变异基因进行功能注释。结果表明:花毛竹基因组检测得到的基因变异数最多,为12 555个; 金丝毛竹样品变异位点数最少,为11 923个; 4个样品都有7 000多个变异基因得到功能注释。GO注释分类包括细胞组件、分子功能和生物过程三个基因功能分类体系的56个功能组。在细胞组件方面,叶绿素合成相关基因有2 431个; 在生物过程方面,参与类胡萝卜素合成过程的基因有75个,参与花青素合成过程中的调控以及紫外光下组织中花青素积累的相关基因有80个。COG分类表明参与复制、重组和修复的基因数为369个,信号转导机制的基因数为291个,转录的相关基因为222个。通过KEGG数据库系统地分析变异基因参与的黄酮类、类胡萝卜素等物质代谢合成途径。深入研究这些差异基因的调控途径,从DNA水平上解释竹秆的变异机制,可为深入研究毛竹种内丰富的多态性和遗传变异提供数据支持,阐析不同变异类型的基因家族、功能基因等遗传基础。     

The association of hormone signalling genes,transcription and changes in shoot anatomy during moso bamboo growth

Moso bamboo is a large, woody bamboo with the highest ecological, economic and cultural value of all the bamboo types and accounts for up to 70% of the total area of bamboo grown. However, the spatiotemporal variation role of moso bamboo shoot during growth period is still unclear. We found that the bamboo shoot growth can be divided into three distinct periods, including winter growth, early growth and late growth based on gene expression and anatomy. In the early growth period, lateral buds germinated from the top of the bamboo joint in the shoot tip. Intercalary meristems grew vigorously during the winter growth period and early growth period, but in the late growth period, mitosis in the intercalary meristems decreased. The expression of cell cycle‐associated genes and the quantity of differentially expressed genes were higher in early growth than those in late growth, appearing to be influenced by hormonal concentrations. Gene expression analysis indicates that hormone signalling genes play key roles in shoot growth, while auxin signalling genes play a central role. In situ hybridization analyses illustrate how auxin signalling genes regulate apical dominance, meristem maintenance and lateral bud development. Our study provides a vivid picture of the dynamic changes in anatomy and gene expression during shoot growth in moso bamboo, and how hormone signalling‐associated genes participate in moso bamboo shoot growth.     

毛竹种群向针阔林扩张的根系形态可塑性

为了弄清毛竹(Phyllostachys edulis)向针阔林扩张过程中根系的形态可塑性反应,在浙江天目山自然保护区毛竹向针阔林扩张的典型过渡地带,连续区域上设置毛竹纯林、针阔-毛竹混交林(以下简称过渡林)、针阔林3种样地。用根钻法采集样地毛竹根系、针阔树根系并比对其生物量密度、细根比根长、相邻同级侧根节点距等形态特征参数变化。结果表明:随着毛竹的扩张程度增加,林内根系生物量密度增加;且与针阔树竞争过程中毛竹将更多的根系放置于表层;同时在水平方向上随离样株距离的增加未出现明显变化,而针阔树根系则随离样木距离的增加而逐渐减少;毛竹根系比根长明显增加,平均增幅15%;一、二级侧根节点距则均有所下降,毛竹侧根数量增多。这些结果表明毛竹种群可通过根系生物量密度、细根比根长、相邻同级侧根节点距等形态可塑性方式实现向周边森林扩张。     

Comparisons of soil‐water content between a Moso bamboo (Phyllostachys pubescens) forest and an evergreen broadleaved forest in western Japan

In Japan, forests of Moso bamboo (Phyllostachys pubescens, an exotic invasive giant bamboo) have naturalized and expanded rapidly, replacing surrounding broadleaved and coniferous forests. To evaluate impacts caused by these forest‐type replacements on the hydrological cycle, soil‐water content and its spatial variability in a Moso bamboo forest were compared with those in an adjacent evergreen broadleaved forest, in a case study of a stand in western Japan (northern Kyushu). The volumetric soil‐water content averaged over depths between 0 and 60 cm was consistently higher in the bamboo stand than that in the broadleaved stand. These results contrast with previous studies comparing the soil‐water content in Moso bamboo forests with that in other forest types. The sum of canopy transpiration and soil evaporation (E) in the bamboo stand tended to be larger than that in the broadleaved stand. Small canopy interception loss was reported in the bamboo forest. Therefore, the large amount of E would counterbalance the small canopy interception loss in the bamboo forest. Differences in soil characteristics between the two stands may be the main factor causing differences in soil‐water content. Spatial variation in soil‐water content in the bamboo stand was larger than that in the broadleaved stand, confirming findings in a previous series of our study. This could happen because the well‐developed root‐system in the bamboo forest enhances preferential flow in the soil. To evaluate the effects of aggressive invasion of alien giant bamboo on the ecosystem functions, we recommend further studies measuring various hydrological components in various Moso bamboo forests.     

C4‐type gene expression is not directly dependent on Kranz anatomy in an amphibious sedge Eleocharis vivipara Link

Eleocharis vivipara Link alters its photosynthetic mode depending on the growth environment. It utilizes C4 photosynthesis when grown under terrestrial conditions (terrestrial form) and C3 photosynthesis when grown under submerged conditions (submerged form). The photosynthetic organ (the mature internodal region of the culm) of the terrestrial form shows typical Kranz anatomy with well-developed bundle sheath cells, while the bundle sheath cells of the submerged form are not developed. In the mature internodal region of the terrestrial form, expression of the genes encoding two carboxylases, the small subunit of ribulose 1,5-bisphosphate carboxylase (RbcS) and phosphoenolpyruvate carboxylase (Ppc), occurred mainly in bundle sheath cells and in mesophyll cells, respectively, as seen in a typical C4 leaf. In the submerged form, RbcS was expressed in both bundle sheath cells and mesophyll cells, and no expression of Ppc was observed. In the immature internodal region with undeveloped bundle sheath cells, both life forms showed the same expression pattern as in C3 plants: RbcS expression was localized in mesophyll cells and no Ppc expression was observed. The C4-type expression pattern was established concomitantly with the development of bundle sheath cells during tissue maturation in the terrestrial internode. In contrast to the terrestrial form, the submerged form maintains C3-type gene expression during tissue maturation. When the terrestrial culm was submerged, a region of transition from the terrestrial form to the submerged form was established in newly sprouting culms. In this transitional region, C4-type expression of the two carboxylase genes was still maintained even though the development of bundle sheath cells was repressed. This observation suggests that the C4-type cell-specific gene expression pattern does not depend on the formation of Kranz anatomy.     

Molecular characterization,phylogenetic analysis and expression profiling of myoglobin and cytoglobin genes in response to heat stress in channel catfish Ictalurus punctatus

To understand the function of myoglobin (Mb) and cytoglobin (Cygb) in channel catfish Ictalurus punctatus in response to heat stress, mb and cygb genes were identified and characterized in this study. These genes were widely expressed in all the tested tissues, but strong tissue preferences were observed, with the mb gene being expressed most highly in the heart, cygb1 most highly expressed in the intestine and cygb2 most highly expressed in the brain. After heat‐stress challenge, mb and cygb genes were up‐regulated in almost all tested tissues. In general, such up‐regulation was more dramatic in the tolerant group than in the intolerant group, suggesting that higher expression of mb and cygb genes contributed to greater tolerance of I. punctatus to heat stress.     

Temporal and spatial profiling of internode elongation-associated protein expression in rapidly growing culms of bamboo

In natural conditions, culms of developing Moso bamboo, Phyllostachys heterocycla var. pubescens, reach their final height of more than ten meters within a short period of two to four months. To study this phenomenon, bamboo culm material collected from different developmental stages and internodes was analyzed. Histological observations indicated that the development of culm was dominated by cell division in the initial stages and by cell elongation in the middle and late stages. Development, maturation, and aging in different regions of the culm were studied systematically from the basal to the top internode. The four major endogenous hormones, indole acetic acid, gibberellic acid, zeatin riboside, and abscisic acid appeared to strongly influence the cell elongation phase. A total of 258 spots were differentially expressed in culm development. Of these, 213 spots were identified by MALDI-TOF/TOF MS and were involved in many physiological and metabolic processes including carbohydrate metabolism, cell division, cell expansion, protein synthesis, amino acid metabolism and redox homeostasis. These proteins with different expression patterns constructed an ingenious network to regulate the culm development. Developmental stage-specific and internode-specific protein expression patterns were identified. Protein abundance was regulated temporally and to some extent spatially, and the sequential development from base to apex of bamboo culm was implemented by temporal and spatial expression of enzymes. Results indicate that during development energy was mainly derived from sucrose degradation, as photosynthetic capacity was poor. The regulation of anaerobic and aerobic modes of respiration appeared to play an important role in energy generation. This is the first report on proteomic profiling in bamboo and helps in understanding the regulatory processes in developing culms.     

酸雨对毛竹入侵阔叶林缓冲区根系形态及分布格局的影响

为明确亚热带毛竹与阔叶树根系对酸雨的适应策略,于2017-2018年在浙江省杭州临安天目山国家级自然保护区毛竹入侵阔叶林过渡区开展酸雨模拟试验,选取T1（pH=4.0）、T2（pH=2.5）、CK（pH=5.5）3个模拟酸雨梯度,分析不同强度酸雨作用下毛竹与阔叶树的根系变化。结果表明：T1对阔叶树根系形态指标具有显著的抑制作用（P<0.05）,导致阔叶树根系的总根长、总表面积、总体积及和细根比根长分别下降了39.9%,39.4%,42.3%和16.2%;T2对毛竹和阔叶树根系指标均有显著的抑制作用（P<0.05）,导致毛竹的根系总根长、总表面积、总体积及细根比根长分别下降了41.5%,42.9%,46.4%和15.1%,阔叶树根系分别下降了60.2%,63.3%,61.8%和20.5%。随着酸雨强度的增加,林内根系生物量密度减少,毛竹将更多的根系置于土壤表层。在水平方向上阔叶树根系则随离树距离的增加而逐渐减少,毛竹2-5 mm径级根系生物量在离竹20 cm,离树40 cm和20 cm之间差异显著（P<0.05）。pH、有机质、可溶有机碳、碱解氮、有效磷和速效钾对毛竹的根系可塑性具有显著的影响（P<0.05）。这些结果表明,毛竹在酸雨作用下利用根系形态可塑性实现种群竞争优势。     

Genome‐wide identification of chitin‐binding proteins and characterization of BmCBP1 in the silkworm,Bombyx mori

The insect cuticle plays important roles in numerous physiological functions to protect the body from invasion of pathogens, physical injury and dehydration. In this report, we conducted a comprehensive genome-wide search for genes encoding proteins with peritrophin A-type (ChtBD2) chitin-binding domain (CBD) in the silkworm, Bombyx mori. One of these genes, which encodes the cuticle protein BmCBP1, was additionally cloned, and its expression and location during the process of development and molting in B. mori were investigated. In total, 46 protein-coding genes were identified in the silkworm genome, including those encoding 15 cuticle proteins analogous to peritrophins with one CBD (CPAP1s), nine cuticle proteins analogous to peritrophins with three CBD (CPAP3s), 15 peritrophic membrane proteins (PMPs), four chitinases, and three chitin deacetylases, which contained at least one ChtBD2 domain. Microarray analysis indicated that CPAP-encoding genes were widely expressed in various tissues, whereas PMP genes were highly expressed in the midgut. Quantitative polymerase chain reaction and western blotting showed that the cuticle protein BmCBP1 was highly expressed in the epidermis and head, particularly during molting and metamorphosis. An immunofluorescence study revealed that chitin co-localized with BmCBP1 at the epidermal surface during molting. Additionally, BmCBP1 was notably up-regulated by 20-hydroxyecdysone treatment. These results provide a genome-level view of the chitin-binding protein in silkworm and suggest that BmCBP1 participates in the formation of the new cuticle during molting.     

Seasonal shifts in giant panda feeding behavior: relationships to bamboo plant part consumption

The giant panda (Ailuropoda melanoleuca) is classified as a carnivore, yet subsists on a diet comprised almost exclusively of bamboo. Wild and captive giant pandas use highly selective foraging behaviors for processing and consuming bamboo. These behaviors are for the first time quantified in captive giant pandas over a 5‐year period of time showing highly specific seasonal trends. Giant panda feeding behavior was recorded using live video observations of two giant pandas housed at the Memphis Zoo from November 2003 to June 2008. Leaf was the primary plant part consumed from June to December, whereas culm was consumed primarily from February to May, with both bears displaying similar seasonal shifts in plant part consumption. From May to June, leaf consumption increased significantly (P‐values<0.001); from June to August, leaf consumption remained high and stable. From December to March, leaf consumption decreased significantly (P‐values<0.001). Specific behaviors for bamboo leaf and culm consumption were also observed. Both bears formed wads of leaves before ingestion while feeding on leaf, but the male employed this feeding behavior more often than the female (54 and 33%, respectively). Both bears used similar culm‐stripping behavior (26 and 25%), used to remove the outer layer and isolate the pith for consumption. This study indicates that unique seasonal foraging behaviors observed in wild pandas are also apparent in captive animals in relation to plant part selectivity and feeding behaviors. Zoo Biol 29:470–483, 2010. © 2009 Wiley‐Liss, Inc.     

Radiocesium Distribution in Bamboo Shoots after the Fukushima Nuclear Accident

The distribution of radiocesium was examined in bamboo shoots, Phyllostachys pubescens, collected from 10 sites located some 41 to 1140 km from the Fukushima Daiichi nuclear power plant, Japan, in the Spring of 2012, 1 year after the Fukushima nuclear accident. Maximum activity concentrations for radiocesium ¹³⁴Cs and ¹³⁷Cs in the edible bamboo shoot parts, 41 km away from the Fukushima Daiichi plant, were in excess of 15.3 and 21.8 kBq/kg (dry weight basis; 1.34 and 1.92 kBq/kg, fresh weight), respectively. In the radiocesium-contaminated samples, the radiocesium activities were higher in the inner tip parts, including the upper edible parts and the apical culm sheath, than in the hardened culm sheath and underground basal parts. The radiocesium/potassium ratios also tended to be higher in the inner tip parts. The radiocesium activities increased with bamboo shoot length in another bamboo species, Phyllostachys bambusoides, suggesting that radiocesium accumulated in the inner tip parts during growth of the shoots.     

Rice plastidial NAD‐dependent malate dehydrogenase 1 negatively regulates salt stress response by reducing the vitamin B6 content

Salinity is an important environmental factor that adversely impacts crop growth and productivity. Malate dehydrogenases (MDHs) catalyse the reversible interconversion of malate and oxaloacetate using NAD(H)/NADP(H) as a cofactor and regulate plant development and abiotic stress tolerance. Vitamin B6 functions as an essential cofactor in enzymatic reactions involved in numerous cellular processes. However, the role of plastidial MDH in rice (Oryza sativa) in salt stress response by altering vitamin B6 content remains unknown. In this study, we identified a new loss‐of‐function osmdh1 mutant displaying salt stress‐tolerant phenotype. The OsMDH1 was expressed in different tissues of rice plants including leaf, leaf sheath, panicle, glume, bud, root and stem and was induced in the presence of NaCl. Transient expression of OsMDH1‐GFP in rice protoplasts showed that OsMDH1 localizes to chloroplast. Transgenic rice plants overexpressing OsMDH1 (OsMDH1OX) displayed a salt stress‐sensitive phenotype. Liquid chromatography–mass spectrometry (LC‐MS) metabolic profiling revealed that the amount of pyridoxine was significantly reduced in OsMDH1OX lines compared with the NIP plants. Moreover, the pyridoxine content was higher in the osmdh1 mutant and lower in OsMDH1OX plants than in the NIP plants under the salt stress, indicating that OsMDH1 negatively regulates salt stress‐induced pyridoxine accumulation. Furthermore, genome‐wide RNA‐sequencing (RNA‐seq) analysis indicated that ectopic expression of OsMDH1 altered the expression level of genes encoding key enzymes of the vitamin B6 biosynthesis pathway, possibly reducing the level of pyridoxine. Together, our results establish a novel, negative regulatory role of OsMDH1 in salt stress tolerance by affecting vitamin B6 content of rice tissues.