甘蓝型油菜分枝角度QTL定位及候选基因分析

分枝角度是油菜重要株型性状.为筛选和发现适度紧凑的分枝角度调控基因,以提高产量利于机械化收获,以分枝角度差异显著的油菜品种Holly和APL01为亲本构建的重组自交系(RIL)群体为材料,在2个环境中对分枝角度进行QTL定位及候选基因分析.结果表明,RIL群体分枝角度表现出连续变异且呈正态分布.利用前期构建的高密度SN...     

Verrucarin A Inhibition of MAP Kinase Activation in a PMA-stimulated Promyelocytic Leukemia Cell Line

Verrucarin A is an inhibitor of protein synthesis. In this study, we examined the inhibitory action of verrucarin A on signal molecules. Verrucarin A partially inhibited the IL-8 production of a PMA-stimulated promyelocytic leukemia cell line (HL-60 cells), and the effect was related to the inhibition of NF-κB activation at non-cytotoxic concentrations. Moreover, the inhibition of mitogen activated protein (MAP) kinase by verrucarin A was especially strong with p38- and JNK-phosphorylation. The findings show a new action of verrucarin A, and it is expected that this action relaxes the signal activation in response to stress.     

Expression of ROR1 in patients with renal cancer--a potential diagnostic marker

Background: The ectopic expression of receptor tyrosine kinase Ror1 has been reported in patients with hematological malignancies such as chronic lymphocytic leukemia and acute lymphoblastic leukemia. Here we report, for the first time, expression of ROR1 gene in both tumor tissues and peripheral blood mononuclear cells (PBMC) from patients with renal cancer (RC). Methods: In the current study, the expression of ROR1 gene was semi-quantitatively measured in PBMC and tumor tissues from 16 RC patients as well as PBMC from 22 healthy individuals relative to the expression of the housekeeping gene phosphoglucomutase 1 by RT-PCR. Results: Our results showed that ROR1 was expressed at gene level in 81.3% of renal tumor tissues (13 out of 16) whereas it was expressed in 94% of PBMC from RC patients (15 out of 16). A weak expression of ROR1 was observed in PBMC of 4 out of 22 healthy individual. A significant expression of ROR1 was observed in PBMC from RC patients when compared to that in PBMC from normal healthy individuals (P<0.001). The expression of ROR1 in PBMC may reflect a shedding of tumor cells into blood stream. Conclusion: We conclude that detection of a high level of ROR1 expression in blood cells might assist in early detection of renal malignancies, providing taking into consideration the clinical symptoms of the disease. Key Words: ROR1, Ectopic expression, Renal cancer     

Exon Sequencing of PKD1 Gene in an Iranian Patient with Autosomal-Dominant Polycystic Kidney Disease

Introduction: Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common genetic kidney disorders with the incidence of 1 in 1,000 births. ADPKD is genetically heterogeneous with two genes identified: PKD1 (16p13.3, 46 exons) and PKD2 (4q21, 15 exons). Eighty five percent of the patients with ADPKD have at least one mutation in the PKD1 gene. Genetic studies have demonstrated an important allelic variability among patients, but very few data are known about the genetic variation among Iranian populations. Methods: In this study, exon direct sequencing of PKD1 was performed in a seven-year old boy with ADPKD and in his parents. The patient’s father was ADPKD who was affected without any kidney dysfunction, and the patient’s mother was congenitally missing one kidney. Results: Molecular genetic testing found a mutation in all three members of this family. It was a missense mutation GTG>ATG at position 3057 in exon 25 of PKD1. On the other hand, two novel missense mutations were reported just in the 7-year-old boy: ACA>GCA found in exon 15 at codon 2241 and CAC>AAC found in exon 38 at codon 3710. For checking the pathogenicity of these mutations, exons 15, 25, and 38 of 50 unrelated normal cases were sequenced. Conclusion: our findings suggested that GTG>ATG is a polymorphism with high frequency (60%) as well as ACA>GCA and CAC>AAC are polymorphisms with frequencies of 14% and 22%, respectively in the population of Southwest Iran. Key Words: Autosomal dominant polycystic kidney disease (ADPKD), Polycystic kidney diseases (PKD), PKD1 gene, Iran     

Smenospongine,a Sesquiterpene Aminoquinone from a Marine Sponge,Induces G1 Arrest or Apoptosis in Different Leukemia Cells

Smenospongine, a sesquiterpene aminoquinone isolated from the marine sponge Dactylospongia elegans, was previously reported by us to induce erythroid differentiation and G1 phase arrest of K562 chronic myelogenous leukemia cells. In this study, we investigated the effect of smenospongine on the cell cycles of other leukemia cells, including HL60 human acute promyelocytic leukemia cells and U937 human histiocytic lymphoma cells by flow cytometric analysis. Smenospongine induced apoptosis dose-dependently in HL60 and U937 cells. The smenospongine treatment increased expression of p21 and inhibited phosphorylation of Rb in K562 cells, suggesting the p21-Rb pathway play an important role in G1 arrest in K562 cells. However, the p21 promoter was not activated by the smenospongine treatment based on a luciferase assay using the transfected K562 cells. Smenospongine might induce p21 expression via another mechanism than transactivation of p21 promoter.     

甘蓝型油菜花瓣缺失性状的主基因+多基因遗传分析

以无花瓣材料APL01和常规四花瓣品种NB6和M083杂交并自交及回交所获得的6个基本世代（P1、P2、F1、B1、B2和F2）为材料，利用主基因+多基因混合遗传模型对无花瓣性状进行遗传分析，结果表明，甘蓝型油菜无花瓣性状的遗传适合E-0模型，即2对加性-显性-上位性主基因+加性-显性-上位性多基因遗传模型。2对主基因的加性效应相等，但不同组合、不同年份的估计值有差异，加性效应值为-11.13 ~ -20.08；2对主基因的显性效应值在不同组合间存在差异，（APL01/NB6）杂交组合估计的2对主基因的显性效应值不同，其中1对基因的效应值较大，而（APL01/M083）估计的2对主基因的显性效应值无差异；上位性效应不同组合间表现也有差异，（APL01/NB6）组合以加加上位和加显上位为主，（APL01/M083）组合以显显上位为主。主基因的遗传率较大，为76.29%-94.13%，不同群体的估计值有差异，以B1世代的估计值较大，B2的估计值较小；多基因的遗传率较小，不同组合、不同年份间表现一致。     

Use of advanced recombinant lines to study the impact and potential of mutations affecting starch synthesis in barley

The effects on barley starch and grain properties of four starch synthesis mutations were studied during the introgression of the mutations from diverse backgrounds into an elite variety. The lys5f (ADPglucose transporter), wax (granule-bound starch synthase), isa1 (debranching enzyme isoamylase 1) and sex6 (starch synthase IIa) mutations were introgressed into NFC Tipple to give mutant and wild-type BC₂F₄ families with different genomic contributions of the donor parent. Comparison of starch and grain properties between the donor parents, the BC₂F₄ families and NFC Tipple allowed the effects of the mutations to be distinguished from genetic background effects. The wax and sex6 mutations had marked effects on starch properties regardless of genetic background. The sex6 mutation conditioned low grain weight and starch content, but the wax mutation did not. The lys5 mutation conditioned low grain weight and starch content, but exceptionally high β-glucan contents. The isa1 mutation promotes synthesis of soluble α-glucan (phytoglycogen). Its introgression into NFC Tipple increased grain weight and total α-glucan content relative to the donor parent, but reduced the ratio of phytoglycogen to starch. This study shows that introgression of mutations into a common, commercial background provides new insights that could not be gained from the donor parent.     

Identification of a Novel Stop Loss Mutation in P2RX2 Gene in an Iranian Family with Autosomal Nonsyndromic Hearing Loss

Background:Hearing loss, a congenital genetic disorder in human, is difficult to diagnose. WES is a powerful approach for ethiological disgnosis of such disorders. Methods:One Iranian family with two patients were attented in the study. Sequencing of known NSHL genes was carried out to recognize the genetic causes of HL. Results:Molecular analyses identified a novel stop loss mutation, c.1048T>G (p.Term350Glu), whitin the P2RX2 gene, causing a termination-site modification.This event would lead to continued translation into the 3'' UTR of the gene, which in turn may result in a longer protein product. The mutation was segregating with the disease phenotype and predicted to be pathogenic by bioinformatic tools. Conclusion:This study is the first Iranian case report of a diagnosis of ADNSHL caused by P2RX2 mutation. The recognition of other causative mutations in P2RX2 gene more supports the probable function of this gene in causing ADNSHL. Key Words: Autosomal dominant 41, Deafness, Mutation, P2RX2, Whole exome sequencing     

Chromosomal changes in uroepithelial carcinomas

This article reviews and summarizes chromosomal changes responsible for the initiation and progression of uroepithelial carcinomas. Characterization of these alterations may lead to a better understanding of the genetic mechanisms and open the door for molecular markers that can be used for better diagnosis and prognosis of the disease. Such information might even help in designing new therapeutic strategies geared towards prevention of tumor recurrences and more aggressive approach in progression-prone cases.The revision of 205 cases of uroepithelial carcinomas reported with abnormal karyotypes showed karyotypic profile characterized by nonrandom chromosomal aberrations varying from one or few changes in low-grade and early stage tumors to massively rearranged karyotypes in muscle invasive ones. In general, the karyotypic profile was dominated by losses of chromosomal material seen as loss of entire chromosome and/or deletions of genetic materials. Rearrangements of chromosome 9 resulting in loss of material from 9p, 9q, or of the entire chromosome were the most frequent cytogenetic alterations, seen in 45% of the cases. Whereas loss of material from chromosome arms 1p, 8p, and 11p, and gains of chromosome 7, and chromosome arm 1q, and 8q seem to be an early, but secondary, changes appearing in superficial and well differentiated tumors, the formation of an isochromosome for 5p and loss of material from 17p are associated with more aggressive tumor phenotypes. Upper urinary tract TCCs have identical karyotypic profile to that of bladder TCCs, indicating the same pathogenetic mechanisms are at work in both locales. Intratumor cytogenetic heterogeneity was not seen except in a few post-radiation uroepithelial carcinomas in which distinct karyotypic and clonal pattern were characterized by massive intratumor heterogeneity (cytogenetic polyclonality) with near-diploid clones and simple balanced and/or unbalanced translocations. In the vast majority of cases strong correlation between the tumors grade/stage and karyotypic complexity was seen, indicating that progressive accumulation of acquired genetic alterations is the driving force behind multistep bladder TCC carcinogenesis. Although most of these cytogenetic alterations have been identified for many years, the molecular consequences and relevant cancer genes of these alterations have not yet been identified. However, loss of TSG(s) from chromosome 9 seems to be the primary and important event(s) in uroepithelial carcinogenesis     

黄顶菊对小麦根尖细胞的遗传毒性

为了探讨入侵植物黄顶菊的遗传毒性,以5个小麦品种为材料,研究5种不同浓度的黄顶菊浸提液对小麦根尖细胞有丝分裂的影响。结果表明,黄顶菊浸提液对小麦有明显的遗传毒性。浸提液抑制了小麦根尖细胞有丝分裂指数,使间期细胞出现微核、多核,使分裂期细胞出现染色体断片、染色体桥、多极分裂、染色体粘连等畸变现象。随着浸提液浓度增加,小麦根尖细胞有丝分裂指数下降,微核率和染色体畸变率升高。     

Development and Identification of Introgression Lines from Cross of Oryza sativa and Oryza minuta

Introgression line population is 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 minuta(accession No.101133) with BBCC genome,as the donor,and an elite indica cultivar IR24(O.sativa),as the recipient.Introgression segments from O.minuta were screened using 164 polymorphic simple sequence repeat(SSR) markers in the genome of each IL.Introgressed segments carried by 131 ILs covered the whole O.sativa genome.The average number of homozygous O.minuta segments per introgression line was about 9.99.The average length of introgressed segments was approximate 14.78 cM,and about 79.64% of these segments had sizes less than 20 cM.In the genome of each introgression line,the O.minuta chromosomal segments harbored chromosomal fragments of O.sativa ranging from 1.15% to 27.6%,with an overall average of 8.57%.At each locus,the ratio of substitution of O.minuta alleles had a range of 1.5% 25.2%,with an average of 8.3%.Based on the evaluation of the phenotype of these ILs,a wide range of alterations in morphological and yield-related traits were found.After inoculation,ILs 41,11 and 7 showed high resistance to bacterial blight,brown planthopper and whitebacked planthopper,respectively.These O.minuta-O.sativa ILs will serve as genetic materials for identifying and using favorable genes from O.minuta.     

Clinical and Genetic Characteristics of Splicing Variant in CYP27A1 in an Iranian Family with Cerebrotendinous xanthomatosis

Background:CTX is a rare congenital lipid-storage disorder, leading to a progressive multisystem disease. CTX with autosomal recessive inheritance is caused by a defect in the CYP27A1 gene. Chronic diarrhea, tendon xanthomas, neurologic impairment, and bilateral cataracts are common symptoms of the disease. Methods:Three affected siblings with an initial diagnosis of non-syndromic intellectual disability were recruited for further molecular investigations. To identify the possible genetic cause(s), WES was performed on the proband. Sanger sequencing was applied to confirm the final variant. The clinical and molecular genetic features of the three siblings from the new CTX family and other patients with the same mutations, as previously reported, were analyzed. The CYP27A1 gene was also studied for the number of pathogenic variants and their location. Results:We found a homozygous splicing mutation, {"type":"entrez-nucleotide","attrs":{"text":"NM_000784","term_id":"1519312912"}}NM_000784: exon6: c.1184+1G>A, in CYP27A1 gene, which was confirmed by Sanger sequencing. Among the detected pathogenic variants, the splice site mutation had the highest prevalence, and the mutations were mostly found in exon 4. Conclusion:This study is the first to report the c.1184+1G>A mutation in Iran. Our findings highlight the other feature of the disease, which is the lack of relationship between phenotype and genotype. Due to nonspecific symptoms and delay in diagnosis, CYP27A1 genetic analysis should be the definitive method for CTX diagnosis. Key Words: Cerebrotendinous xanthomatosis, CYP27A1, Intellectual disability, Iran, Whole exome sequencing     

甘蓝型油菜和甘蓝CRISPR/Cas9编辑效果的快速检测

CRISPR/Cas9系统是目前最常用的基因组定点编辑工具，通过瞬时表达试验提前验证Cas9/sgRNA载体诱导的突变效率，可以提高基因组定点编辑成功的几率，且显著节省费用及时间。本研究开发了一种利用农杆菌介导的操作简单、适用性好、成本低廉的叶片瞬时表达技术，可用于快速检测甘蓝型油菜和甘蓝中CRISPR/Cas9的编辑效果。针对甘蓝型油菜BnaC.WRKY11.a设计了2个靶位点Tgt1（Target 1）和Tgt2（Target 2），并构建了Cas9/sgRNA-Tgt1/2多重突变载体，在甘蓝型油菜叶片中瞬时表达后，2个靶位点都出现突变，突变效率达11.2%～82.2%。针对甘蓝BolPDS3基因设计了1个靶位点Tgt3（Target3），并构建了Cas9/sgRNA-Tgt3敲除载体，在甘蓝叶片瞬时表达后，Tgt3发生了突变，且大部分为碱基缺失突变，缺失的数目为1～18bp不等，同时还存在少量碱基插入以及碱基替换等突变类型。结果表明这种甘蓝型油菜和甘蓝的叶片瞬时表达技术操作简单、适用性好、成本低廉，CRISPR/Cas9的编辑效果快速易检测。     

20个云南无性系茶树良种的DNA指纹图谱构建

以20个云南无性系茶树良种为材料,选用7对多态性丰富、品种区分率高、易统计的ISSR引物对茶树良种进行分析。结果显示,7对ISSR引物共扩增出110条带,其中多态带93个,多态条带比例为84.54%,多态信息量平均为0.417,品种相似性系数在0.574~0.854之间,其中引物UBC835,ISSR2不仅多态性丰富,且单个引物就可区分所有品种,是最有效的核心引物;7对核心引物两两组合的效率分析表明,UBC835/UBC811、ISSR2/UBC835和UBC835/ISSR3是高效引物组合,可以完全有效区分所有品种,且品种相似性系数较低;同时使用15个地方品种验证3个高效引物组合,结果表明,UBC835/ISSR2是最佳引物组合,不仅能有效区分所有云南无性系茶树良种,而且能将云南无性系良种与15个地方品种最有效地区分开。使用品种的国家地区代码、育种单位英文缩写、核心引物名称和分子数据组成云南无性系茶树良种的DNA指纹图谱,不仅包含了品种的重要信息,而且其中的分子数据可用作茶树品种的真伪鉴定和遗传关系分析,为茶树品种的知识产权保护提供有效的科学依据。     

分子标记技术在小麦遗传育种中的应用现状

小麦庞大的基因组使得分子标记技术在小麦中的应用落后于大麦、玉米、水稻等作物。近年来,随着他子标记技术检测系统的发展和完善,分子标记技术在小麦中的应用已有了很大的进展。分子标记技术依靠提供准确、稳定可靠的ＤＮＡ水平的遗传标记,在小麦遗传育种研究中已用于构建遗传图谱、标定和定位目的基因、鉴定与标记外源染色体片段、鉴定品种真实性和纯度、绘制品种指纹图谱、遗传分析、物种演变、标记辅助选育等方面。     

大麦EMS突变体库构建和突变体筛选

为研究大麦重要功能基因并给大麦遗传育种提供新的种质资源,通过甲基磺酰乙酯(EMS)诱变处理优质、高抗黄花叶病大麦品种苏啤6号,从2400个M_2代株系中筛选出152个突变株系,突变频率为6.33%。其中,幼苗习性突变株系36个,突变率为1.50%;叶部突变株系34个,突变率为1.42%;茎部突变株系33个,突变体率为1.38%;穗部突变株系22个,突变率为0.92%;成熟期和育性突变株系27个,突变率为1.13%。经过M_3代验证,获得可稳定遗传的各类突变株系43个。     

EMS诱导的盛农1号小麦突变体筛选与鉴定

突变体诱导是创造小麦遗传材料和选育新品种的重要途径之一。通过甲基磺酸乙酯（EMS）处理高产、抗白粉病品系盛农1号,从4 500个M₂代穗系中筛选出174个突变体穗系,突变率为3.86%。其中,穗部突变体32个,突变率为0.71%;叶部突变体47个,突变率为0.89%;茎部突变体22个,突变率为0.49%;育性和成熟期突变体12个,突变率为0.27%;其他类型突变体11个,突变率为0.24%;农艺性状优于对照盛农1号的突变体57个,突变率为1.27%。经过M₃代验证及生物学性状和农艺性状调查,获得可稳定遗传的株系18个,其中,穗部突变体6个,茎秆突变体1个,叶部突变体6个,早熟突变体3个,晚熟突变体2个。农艺性状分析表明,穗部突变体SS4、SS5、SS6和SS16的小穗性状优于盛农1号。这些新突变体为小麦品种改良和遗传研究提供了新的材料。     

利用CRISPR/Cas9系统定向改良水稻稻瘟病抗性

【目的】CRISPR/Cas9基因编辑技术是作物遗传改良的有效工具。本研究通过对水稻Pita、Pi21和ERF922稻瘟病相关基因进行定点编辑，以期获得能够稳定遗传的抗稻瘟病水稻材料。【方法】利用CRISPR/Cas9基因编辑技术，以Pita、Pi21和ERF922为靶基因，构建共编辑载体pC1300-2×35S::Cas9-gPita-gPi21-gERF922 ，用农杆菌转化长粒粳稻恢复系L1014，筛选获得稳定遗传的纯合突变体用于稻瘟病抗性鉴定。【结果】在T0代转基因株系中，Pita、Pi21和ERF922突变频率分别为75%、85%和65%，突变基因型多为双等位突变。筛选到的不含T-DNA成分的T1代能够稳定遗传给T2代，并从中获得Pi21单突变纯合株系及Pita、Pi21和ERF922的三突变纯合株系。稻瘟病抗性鉴定结果表明，与野生型相比，突变株系的抗性显著提高。同时，接种后纯合突变体株系内水杨酸、茉莉酸和乙烯等信号转导途径相关基因的表达量均上调。据此，我们推测纯合突变株系对稻瘟病的抗性增强可能与其对稻瘟病菌的响应被激活有关。【结论】利用CRISPR/Cas9技术获得了能够稳定遗传和具有较高稻瘟病抗性的纯合突变株系，为水稻稻瘟病抗性改良提供了良好的材料。     

利用CRISPR/Cas9系统定向改良水稻稻瘟病抗性

[目的]CRISPR/Cas9 基因编辑技术是作物遗传改良的有效工具。本研究通过对水稻Pita、Pi21和ERF922稻瘟病相关基因进行定点编辑,以期获得能够稳定遗传的抗稻瘟病水稻材料。[方法]利用 CRISPR/Cas9基因编辑技术,以 Pita、Pi21 和 ERF922 为靶基因,构建共编辑载体 pC1300-2×35S::Cas9-g^Pita-g^Pi21-g^ERF922 ,用农杆菌转化长粒粳稻恢复系L1014,筛选获得稳定遗传的纯合突变体用于稻瘟病抗性鉴定。[结果]在T0代转基因株系中,Pita、Pi21和ERF922 突变频率分别为 75%、85%和 65%,突变基因型多为双等位突变。筛选到的不含 T-DNA成分的T1代能够稳定遗传给T2代,并从中获得 Pi21单突变纯合株系及Pita、Pi21和ERF922的三突变纯合株系。稻瘟病抗性鉴定结果表明,与野生型相比,突变株系的抗性显著提高。同时,接种后纯合突变体株系内水杨酸、茉莉酸和乙烯等信号转导途径相关基因的表达量均上调。据此,我们推测纯合突变株系对稻瘟病的抗性增强可能与其对稻瘟病菌的响应被激活有关。[结论]利用 CRISPR/Cas9 技术获得了能够稳定遗传和具有较高稻瘟病抗性的纯合突变株系,为水稻稻瘟病抗性改良提供了良好的材料。