基于转录组测序的高羊茅分蘖与株高相关差异表达基因分析

分蘖与株高是禾本科草类植物重要的农艺性状,明确参与调控分蘖与株高的基因类型对牧草和草坪草分子辅助育种具有重要意义。以表型差异明显的两个高羊茅品种“Kentucky-31”（K31）和“Regenerate”为材料,旨在构建高羊茅分蘖节转录组图谱,挖掘在分蘖节部位调控生长发育相关的差异表达基因（differentially expressed genes,DEGs）。基于高通量测序技术平台Illumina HiSeq 2500×Miseq 300进行转录组测序,并将得到的数据进行de novo组装,结果共获得77872条单基因簇（unigene）。将获得的unigenes与非冗余蛋白数据库（non-redundant protein database,NR）、蛋白质数据库（universal protein,Uniprot）、基因本体数据库（gene ontology,GO）、东京基因与基因组数据库（kyoto encyclopedia of genes and genomes,KEGG）以及直系同源蛋白簇（clusters of orthologous groups,COG）数据库进行比对,结果显示：分别有59927、40213、44447、15146和13767条unigenes成功获得注释。“Regenerate”与“K31”对比有1573个上调DEGs和1441个下调DEGs。GO富集分析发现,DEGs主要富集在细胞、细胞组分、大分子复合物组装等生物过程。DEGs中共注释到42个差异表达转录因子,主要包括TCP、WRKY和ARF等19种类型。还注释到与8类植物激素相关的DEGs,包括生长素、细胞分裂素、脱落酸、赤霉素、乙烯、油菜素甾醇、水杨酸和茉莉酸。利用实时荧光定量PCR对DEGs进行表达模式验证,发现其与RNA-Seq测序结果一致,证实了测序结果的准确性。研究结果丰富了高羊茅的转录组序列资源,初步获得控制株高及分蘖发育的候选因子,为进一步开展基因功能及分子育种研究提供了理论支持。     

银杏种子生长、脱落及其与内源激素含量变化的关系

关于激素在果实生长发育过程中的作用 ,前人曾进行过大量的研究 ,并取得了许多宝贵的成果[1,2 ] ,而对银杏 (ＧｉｎｋｇｏｂｉｌｏｂａＬ .)种子的相关研究却很少。银杏作为一种古老的裸子植物 ,其种子的生长发育与一般的裸子植物又有不同 ,它的胚珠直接着生在短枝上 ,没有了一般裸子植物的球果构造 ,从表面上看很类似于杏 (Ａｒｍｅｎｉａｃａｓｐｐ .)、桃 (Ａｍｙｇｄａｌｕｓｓｐｐ .)等的生长发育过程。本文从分析银杏种子生长发育的一般特点出发 ,研究了银杏种子生长发育与内源激素含量的关系 ,旨在探索银杏种子生长发育的一般规律 ,…     

小叶杨原生质体培养中的激素和氨含量变化

以小叶杨（Ｐｏｐｕｌｕｓｓｉｍｏｎｉｉｃａｒｒ．）原生质体培养的愈伤组织为供试材料，用高压波相色谱法（ＨＰＬＣ），测定它们在四种不同分化培养基上四大类内源激素（ＩＡＡ，ＧＡ３，ＣＴＫ，ＡＢＡ）的含量变化，并检定有关Ｎ的生化反应。结果表明，不同分化培养基上的愈伤组织中内源激素含量和种类差异很大。导致细胞分裂素（ＣＴＫ）含量高，种类多以及细胞分裂素与生长素的比值（ＣＴＫ／ＩＡＡ）高的处理，有利于愈伤组织分化；但若愈伤组织中含有内源脱落酸（ＡＢＡ），则抑制分化。测定结果还表明，小叶杨原生质体培养的愈伤组织分化过程中，其硝酸还原酶活力与氨基态Ｎ含量互为消长。分化频率高的愈伤组织中，硝酸还原酶活力低，而氨基态Ｎ含量高．     

Content changes of several endogenous plant hormones in female-sterile <Emphasis Type="Italic">Pinus tabulaeformis</Emphasis> Carr

To determine the relationship between female sterility of Pinus tabulaeformis Carr. and plant hormones, content changes of several endogenous plant hormones in a female-sterile clone and a normal clone were analyzed during the key period of female gametophyte abortion in mutant clones. The change of ABA content showed a similar pattern between normal and mutant clones, but the ABA content of the mutant clone was always much higher than that of the normal clone. In the normal clone, the IAA content decreased significantly at the early stage of the period and reached its minimum during the first ten days of April and stabilized thereafter. The endogenous ZR levels increased sharply at the early stages and reached a maximum in the middle of April, then decreased dramatically. The IAA and ZR contents of the mutant clone did not change perceptively through the entire period. We conclude that the high levels of ABA and the deficit of IAA and ZR may be the reasons of female gametophyte abortion in the female-sterile clones of P. tabulaeformis.     

AgNO3处理下银杏幼胚愈伤组织的形成和激素变化

采用石蜡切片技术和酶联免疫吸附法(ELISA),以银杏幼胚为外植体,添加0,1和5 mg/L AgNO3作处理,观察21 d内愈伤组织的形成过程,测定内源激素吲哚乙酸(IAA)、保幼激素类似物(ZR)、赤霉素(GA)和脱落酸(ABA)的变化,分析AgNO3对银杏幼胚愈伤组织分化潜能的影响。结果表明:根据解剖结构愈伤形成可分为4个阶段。第1阶段(0~7 d),IAA升高,ABA降低,表皮细胞启动脱分化。第2阶段(≥7~11 d),4种激素浓度均升高,愈伤组织和细胞团初步形成。第3阶段(≥11~16 d),GA、ABA大幅降低,细胞团大量形成。第4阶段(≥16~21 d),ABA骤降、GA迅速上升,愈伤组织和细胞团增殖。1 mg/L AgNO3处理下分生细胞团种类最多,其次为5mg/L处理,0 mg/L处理最少。研究认为,银杏幼胚愈伤组织具有较强的分化潜能,AgNO3能影响其内源激素的平衡,调控分化方向。     

THYROID SCINTIGRAPHY FINDINGS IN 2096 CATS WITH HYPERTHYROIDISM

Thyroid scintigraphy is currently the reference standard for diagnosing and staging cats with hyperthyroidism, but few studies describing the scintigraphic characteristics in a large number of cats have been reported. The objective of this study was to better characterize thyroid scintigraphy findings by evaluating 2096 consecutive cats with hyperthyroidism that were referred over a 3.5‐year period. Of these cats, 2068 (98.7%) had a high thyroid‐to‐salivary ratio (>1.5), whereas 2014 (96.1%) were found to have a high thyroid‐to‐background ratio (>6.1). When the patterns of the cats’ thyroid disease were recorded, 665 (31.7%) had unilateral disease, 1060 (50.6%) had bilateral‐asymmetric disease (two thyroid lobes unequal in size), 257 (12.3%) had bilateral‐symmetric disease (both lobes similar in size), and 81 (3.9%) had multifocal disease (≥3 areas of increased radionuclide uptake). The number of areas of ^99mTcO^?₄ uptake in the 2096 cats ranged from 1 to 6 (median, 2), located in the cervical area in 2057 (98.1%), thoracic inlet in 282 (13.5%), and in the thoracic cavity in 115 (5.5%). Ectopic thyroid tissue (e.g. lingual or mediastinal) was diagnosed in 81 (3.9%) cats, whereas thyroid carcinoma was suspected in 35 (1.7%) of the cats. The results of this study support conclusions that most hyperthyroid cats have unilateral or bilateral thyroid nodules, but that multifocal disease will develop in a few cats that have ectopic thyroid disease or thyroid carcinoma. Both ectopic thyroid disease and thyroid carcinoma are relatively uncommon in hyperthyroid cats, with a respective prevalence of ～4% and ～2% in this study.     

STRAIN ELASTOGRAPHY USING DOBUTAMINE‐INDUCED CAROTID ARTERY PULSATION IN CANINE THYROID GLAND

Thyroid disease is common in dogs and conventional ultrasonography is a standard diagnostic test for diagnosis and treatment planning. Strain elastography can provide additional information about tissue stiffness noninvasively after applying external or internal compression. However, natural carotid artery pulsations in the canine thyroid gland are too weak to maintain sufficient internal compression force. The objective of the present study was to describe the feasibility of strain elastography for evaluating the canine thyroid gland and the repeatability of dobutamine‐induced carotid artery pulsation as an internal compression method. In seven healthy Beagle dogs, strain on each thyroid lobe was induced by external compression using the ultrasound probe and internal compression using carotid artery pulsation after dobutamine infusion. The thyroid appeared homogeneously green and the subcutaneous fat superficial to the thyroid lobe appeared blue. Strain values and strain ratios did not differ among dogs or between the left and right lobes. Interobserver repeatability was excellent for both compression methods. Intraobserver repeatability of the strain ratio measured using the carotid artery pulsation method (intraclass coefficient correlation = 0.933) was higher than that measured using the external compression method (0.760). Mean strain values of thyroid lobes for the external compression method (142.93 ± 6.67) differed from the internal method (147.31 ± 8.24; P < 0.05). Strain ratios between the two methods did not differ. Strain elastography was feasible for estimating thyroid stiffness in dogs. Carotid artery pulsation induced by dobutamine infusion can be used for canine thyroid strain elastography with excellent repeatability.     

梅PmARF17克隆及其在花发育中与内源激素的调控模式

【目的】分析梅PmARF17的生物学功能,探究梅花发育进程中其表达丰度与内源激素动态变化的关系,为梅花发育的调控研究提供依据。【方法】以梅品种‘大嵌蒂’为试材,克隆PmARF17,利用生物信息学软件分析基因结构、系统进化及其与其他物种同源蛋白的差异;亚细胞定位确定PmARF17蛋白在细胞中作用的部位;以梅品种‘大嵌蒂’和‘龙眼’不同发育阶段的花芽、叶芽、花器官为试材,利用qRT-PCR检测PmARF17时空表达模式,通过UPLC法测定IAA、GA₃、ABA、ZT含量的动态变化,并与PmARF17的表达进行相关性分析;克隆PmARF17启动子,分析启动子的顺式作用元件,利用瞬时表达解析PmARF17与GA₃的调控模式。【结果】从梅品种‘大嵌蒂’中克隆得到PmARF17,系统进化树分析表明PmARF17蛋白与其他植物的ARF蛋白序列高度同源;亚细胞定位表明其作用于细胞核和细胞膜上;qRT-PCR表达和内源激素含量的相关性分析表明,PmARF17的表达与IAA含量的变化趋势没有明显的相关性。PmARF17在雌蕊完好花芽中的表达水平相对不完全花芽显著上调,而GA₃含量与PmARF17的表达趋势一致。ABA和ZT含量总体上与PmARF17的表达呈相反的趋势,表明两者可能抑制PmARF17的表达。PmARF17启动子含有GA顺式元件,且具有启动活性和组织表达特异性,在花瓣、雄蕊及根部特异表达。【结论】 PmARF17可能是梅花发育的正调控基因,促进梅雌蕊的正常发育。PmARF17的表达可能受到GA₃的正调控,其可能通过作用于雄蕊和花瓣,进而影响梅的雌蕊发育进程。     

Molecular cloning of the cDNA encoding the β subunit of thyrotropin and regulation of its gene expression by thyroid hormones in the goldfish, Carassius auratus

A cDNA encoding the subunit of thyrotropin (TSH) was isolated from a goldfish (Carassius auratus) pituitary gland cDNA library. By comparing the sequence with other teleost TSHs, a signal peptide of 19 amino acids and a mature hormone of 131 amino acids were predicted for goldfish TSH subunits. The resulting putative mature hormone of 131 amino acids had well-conserved cysteine positions and a putative N-linked glycosylation site; homology was 51–67% with TSHs from other teleosts, 38–43% with tetrapod TSHs, but only 27 and 29% with goldfish GTH-I and -II, respectively. We also examined the effects of thyroid hormones (TH) and thiourea (TU, an inhibitor of TH production) treatments on TSH and GTH subunit gene expressions in the goldfish pituitary gland. After thyroxine (T₄) treatment, circulating T₄ concentration increased and TSH mRNA level decreased. Supressing the amount of circulating T₄ and triiodothyronine (T₃) by TU treatment increased the TSH mRNA level. Moreover, T₄ replacement therapy (simultaneous treatment of both TU and T₄) caused a high level of circulating T₄ and a low level of circulating T₃, and a decrease in the TSH mRNA level. Thus, changing levels of circulating TH exert a negative feedback on the level of TSH subunit mRNA in goldfish in vivo. On the other hand, GTH subunit mRNA levels were not affected by changes in the levels of circulating TH.     

Low protein provision during the first year of life,but not during foetal life,affects metabolic traits,organ mass development and growth in male mink (Neovison vison)

Low protein provision in utero and post‐partum may induce metabolic disorders in adulthood. Studies in mink have mainly focused on short‐term consequences of low protein provision in utero whereas the long‐term responses to low protein (LP) provision in metabolically programmed mink are unknown. We investigated whether low protein provision in utero affects the long‐term response to adequate (AP) or LP provision after weaning in male mink. Eighty‐six male mink were exposed to low (19% of ME from CP; crude protein) or adequate (31% of ME from CP) protein provision in utero, and to LP (~20% of ME from CP) or AP (30–42% of ME from CP) provision post‐weaning. Being metabolically programmed by low protein provision in utero did not affect the response to post‐weaning diets. Dietary protein content in the LP feed after weaning was below requirements; evidenced by lower nitrogen retention (p < 0.001) preventing LP mink from attaining their growth potential (p < 0.02). LP mink had a lower liver, pancreas and kidney weight (p < 0.05) as well as lower plasma IGF‐1 concentrations at 8 and 25 (p < 0.05) weeks, and a higher incidence of hepatic lipidosis at 25 weeks (p < 0.05). Furthermore, LP mink had a higher body fat (p < 0.05) and lower body CP content (p < 0.05) at 50 weeks of age. It is concluded that some effects of low protein provision in utero can be alleviated by an adequate nutrient supply post‐partum. However, long‐term exposure to low protein provision in mink reduces their growth potential and induces transient hepatic lipidosis and modified body composition.