Base editing with high efficiency in allotetraploid oilseed rape by A3A‐PBE system

CRISPR/Cas‐base editing is an emerging technology that could convert a nucleotide to another type at the target site. In this study, A3A‐PBE system consisting of human A3A cytidine deaminase fused with a Cas9 nickase and uracil glycosylase inhibitor was established and developed in allotetraploid Brassica napus. We designed three sgRNAs to target ALS, RGA and IAA7 genes, respectively. Base‐editing efficiency was demonstrated to be more than 20% for all the three target genes. Target sequencing results revealed that the editing window ranged from C1 to C10 of the PAM sequence. Base‐edited plants of ALS conferred high herbicide resistance, while base‐edited plants of RGA or IAA7 exhibited decreased plant height. All the base editing could be genetically inherited from T₀ to T₁ generation. Several Indel mutations were confirmed at the target sites for all the three sgRNAs. Furthermore, though no C to T substitution was detected at the most potential off‐target sites, large‐scale SNP variations were determined through whole‐genome sequencing between some base‐edited and wild‐type plants. These results revealed that A3A‐PBE base‐editing system could effectively convert C to T substitution with high‐editing efficiency and broadened editing window in oilseed rape. Mutants for ALS, IAA7 and RGA genes could be potentially applied to confer herbicide resistance for weed control or with better plant architecture suitable for mechanic harvesting.     

金针菇高表达基因的密码子偏好性及特征分析

为探究金针菇的密码子偏好性,挖掘高表达基因的特征信息,以金针菇基因组及转录组数据为材料,分析金针菇的密码子偏好性及其影响因素,并对发育阶段高表达基因进行功能注释和顺式元件分析。分析表明,金针菇高表达基因表现出较强的密码子偏好性,且其偏好密码子多以胞嘧啶(C)结尾,此外在高表达基因中存在6种氨基酸的最优密码子较为保守。在进化过程中,金针菇高表达基因密码子偏好性受到自然选择压力的影响较大。功能注释分类表明,高表达基因多为核糖体通路相关的基因,与蛋白质翻译和生物合成相关。顺式元件分析表明,高表达基因启动子区域大多存在MeJA响应元件、ABA响应元件、光响应元件及MYB转录因子结合元件。研究结果可为提高金针菇异源表达效率和挖掘强启动子提供理论基础和思路。     

滇东南普阳盆地含煤地层孢粉组合特征及其年代

云南地处青藏高原东南缘, 生物多样性丰富, 季风气候特征明显。新生代剧烈的构造活动形成了众多山间盆地, 其间保存了大量精美的植物化石, 是探讨新生代以来植被、植物多样性和地球环境演变的理想地区。但是长期以来, 由于地层年代学证据的缺乏, 这些新生代沉积盆地的地质年代还存在很大争议。本研究通过对滇东南地区富宁县普阳盆地的含煤地层开展深入的孢粉学研究, 探讨盆地含煤地层年代及其古气候演化过程。剖面下部煤层孢粉组合以杉粉属(Taxodiaceaepollenites)为主, 指示以杉科为主的湿润沼泽森林, 冬春季相对湿润; 剖面上部孢粉组合指示以常绿栎类为主的亚热带常绿‒落叶阔叶混交林, 生长山核桃粉属(Caryapollenites)、冬青粉属(Ilexpollenites)、胡桃粉属(Juglanspollenites)、枫香粉属(Liquidambarpollenites)等亚热带常见树种, 气候温暖湿润, 季节分明。结合新发现的哺乳动物化石证据和周边地区不同地质时代的孢粉组合, 普阳盆地含煤地层的沉积时代应为晚始新世; 同时, 孢粉组合也表明滇东南地区植被现代化面貌至少在晚始新世就已经开始出现。     

Estrogen receptor beta promotes lung cancer invasion via increasing CXCR4 expression

Lung cancer is one of the most lethal malignant tumors in the world. The high recurrence and mortality rate make it urgent for scientists and clinicians to find new targets for better treatment of lung cancer. Early studies indicated that estrogen receptor β (ERβ) might impact the progression of non-small-cell lung cancer (NSCLC). However, the detailed mechanisms, especially its linkage to the CXCR4-mediated cell invasion, remain unclear. Here we found that ERβ could promote NSCLC cell invasion via increasing the circular RNA (circRNA), circ-TMX4, expression via directly binding to the 5′ promoter region of its host gene TMX4. ERβ-promoted circ-TMX4 could then sponge and inhibit the micro RNA (miRNA, miR), miR-622, expression, which can then result in increasing the CXCR4 messenger RNA translation via a reduced miRNA binding to its 3′ untranslated region (3′UTR). The preclinical study using an in vivo mouse model with orthotopic xenografts of NSCLC cells confirmed the in vitro data, and the human NSCLC database analysis and tissue staining also confirmed the linkage of ERβ/miR-622/CXCR4 signaling to the NSCLC progression. Together, our findings suggest that ERβ can promote NSCLC cell invasion via altering the ERβ/circ-TMX4/miR-622/CXCR4 signaling, and targeting this newly circ-TMX4/miR-622/CXCR4 signaling may help us find new treatment strategies to better suppress NSCLC progression.Subject terms: Non-small-cell lung cancer, Metastasis     

一氧化氮参与盐胁迫下长春花酚类代谢的调控研究

为研究外源一氧化氮(NO)调控盐胁迫下长春花中酚类化合物的响应,采用液相色谱—质谱联用(LCMS)技术靶向分析梯度浓度硝普纳(SNP)处理对盐胁迫下长春花幼苗根、茎、花、叶4个部位中酚类化合物组分及含量水平的变化。结果共鉴定出L-苯丙氨酸和18种酚类物质,C6C1类5种、C6C3类5种、C6C3C6类8种,其中原儿茶酸、绿原酸、槲皮素在长春花根、茎、花、叶4个部位中均存在;不同浓度SNP处理后长春花不同部位酚类化合物响应积累明显不同,其中C6C1和C6C3小分子酚酸类化合物主要积累在根和茎中,C3C6C3类主要富集在花和叶中;L-苯丙氨酸在茎、叶中相对含量较高,盐胁迫下茎中含量显著升高,且随外源NO浓度增大呈下降趋势。外源NO影响盐胁迫下植物器官中酚类化合物的积累和变化,其中根和茎响应敏感,从种类和相对含量的角度,茎和叶更适合检测酚类化合物。     

普氏蹄蝠下丘谐波主频神经元的时程选择性

为探讨下丘(Inferior colliculus,IC)回声定位信号主频范围内的神经元的时程选择性,在自由声场刺激条件下,我们在4 只普氏蹄蝠的IC 采用不同时程的声刺激,研究了神经元的时程选择性。通过在体细胞外记录,共获得56 个声敏感下丘神经元,其记录深度、最佳频率和最小阈值的范围分别为1547 - 3967 (2878. 9 ±629.1)μm,20 -68 (49.0 ± 11. 1)kHz 和36.5 -95. 5 (59. 8 ±13. 0)dB SPL。根据所记录到的下丘神经元对不同时程的声刺激的反应,即对不同时程的选择性(Duration selectivity),将其分为6 种类型:短通型(Short-pass,SP,n = 11/56)、带通型(Band-pass,BP,n = 1/56)、长通型(Long-pass,LP,n = 5 /56)、反带通型(Band-reject,BR,n = 3 /56)、多峰型(Multi-peak,MP,n =6 /56)和全通型(All-pass,AP,n =30 /56)或非时程选择型(Nonduration-selective,NDS)。通过比较普氏蹄蝠下丘谐波主频内和主频外神经元的时程选择性,我们发现处于回声定位信号主频范围内神经元(n =32)比主频外神经元(n = 24)具有更短的最佳时程和更高的时程选择性。结果提示,在普氏蹄蝠回声定位过程中谐波主频内神经元较谐波主频外神经元发挥了更为重要的作用。     

宁夏黄土丘陵区不同生态恢复生境地表甲虫多样性

为了解宁夏黄土丘陵区不同生境地表甲虫群落多样性变化规律及与环境因子的关系, 并探讨不同生态恢复措施对维持地表甲虫群落多样性的影响, 2013年7-8月, 作者利用陷阱法调查了该区6种生境内的地表甲虫群落多样性。结果表明: 灌草混交林地、乔灌混交林地、生态薪炭林地中地表甲虫物种丰富度和个体数量均较高, 天然封育草地、生态经济林地和水平农田中物种丰富度较低, 生态经济林地和水平农田中甲虫个体数量显著高于天然封育草地。不同生境间, 灌草混交林地、乔灌混交林地与生态薪炭林地之间、生态经济林地与水平农田之间甲虫群落组成相似性较高。多元回归分析表明, 草本层生物量、灌木层盖度及土壤含水量是影响甲虫物种丰富度的决定因素, 林冠层盖度和枯落物厚度是决定地表甲虫个体数量的重要因素。CCA分析表明, 枯落物盖度、枯落物厚度、林冠层盖度及草本层盖度是影响地表甲虫群落组成的重要环境因子。研究表明, 灌草混交林地为地表甲虫群落多样性维持较好的生境类型, 是宁夏黄土丘陵区典型生态恢复的最优模式。     

DCs sensitized with mPD-L1-Ig fusion protein improve the effect of heart transplantation in mice by promoting the generation of T-reg cells

Purpose

To detect the effects of DCs sensitized by mPD-L1-Ig fusion protein in heart transplantation in mice as well as its mechanisms.

Method

The mPD-L1-IgG1 construct was used to build a yeast expression system, and the fusion protein was expressed by secretion after the transfection of the GS115 yeast strain, purified by affinity chromatography and ion exchange chromatography, and assayed by SDS–PAGE and Western blot. The ability of the fusion protein to bind to the acceptor PD-1 was tested by ELISA, and the ability of the fusion protein to inhibit the function of T cells was tested by mixed lymphocyte reaction (MLR).

Results

We used the new PD-L1-IgG1 fusion protein to sensitize imDCs and maintained the immature state of DCs, so as to induce stable and effective immune tolerance to heart transplantation. After the treatment of DCs by mPD-L1-Ig in vitro, the levels of CD80, CD40 and I-Ab expression on DCs are relatively weaker, the ability of DCs to stimulates the proliferation of allogeneic spleen T cells was significantly decreased (P < 0.01), and the levels of Th1 (IL-2, IFN-γ) and Th2 (IL-4, IL-10) secreted by induced allogeneic T cells were significantly decreased (P < 0.01). An in vivo experiment also revealed that DCs sensitized by mPD-L1-IgG1 could prolong the survival time of a transplanted heart to 17.8 ± 1.12 days, and alleviate the pathological change of the cardiac allografts compared with other three groups.

Conclusion

DCs sensitized by the yeast-expressed mPD-L1-Ig fusion protein are shown to alleviate the cardiac allograft rejection in mice.     

香蕉MaASR1基因的抗干旱作用

ASR(ABA, stress, ripening induced protein)是一类响应植物干旱胁迫的关键转录因子, 在许多植物中已有报道, 然而尚未见香蕉(Musa acuminata)中ASR与抗旱作用的相关研究。该实验从香蕉果实cDNA文库中筛选出1个ASR基因, 即MaASR1(登录号为AY628102)。干旱胁迫下, 该基因在叶片中的表达量高于根部。将MaASR1转入拟南芥(Arabidopsis thaliana), Southern检测确定了两株独立表达的转基因株系(命名为L14和L38)。表型观察发现, 此两转基因株系的叶片变小且变厚; Northern和Western检测结果表明, MaASR1在L14和L38中表达。控水处理后, L14和L38的存活率及脯氨酸含量均高于野生型。经干旱胁迫和外源ABA处理后, 对MaASR1转基因株系中ABA/胁迫响应基因的表达分析, 发现MaASR1可增强转基因株系对ABA信号的敏感度, 但不能增强植株依赖于ABA途径的抗旱性。     

巨膜长蝽的形态特征和生物学特性

【目的】近年来巨膜长蝽Jakowleffia setulosa(Jakovlev)由以往稳定种群的荒漠昆虫上升为暴发性发生并迁移至农区危害的农业害虫。由于缺乏对巨膜长蝽基础生物学的相关研究,对其开展监测预警和农业防治存在困难。开展巨膜长蝽个体生物学研究,填补研究空白对于有效监测预报和防控提供可靠的依据,同时能为下一步分析巨膜长蝽成灾规律提供基础数据。【方法】本文通过室内观察和野外调查相结合,系统阐明该种形态特征、生活史、习性和行为等生物学特性。【结果】巨膜长蝽在宁夏中部干旱带一年发生2代,5月中旬和10月中下旬是两个发生高峰期,6月中旬至8月中旬以成虫进入滞育状态,11月下旬以成虫越冬;雌成虫的平均寿命为(32.14±2.34)d,雄成虫的平均寿命为(28.00±3.13)d,雌雄性比为1︰1.9,有多次交尾多次产卵的习性,每头雌虫抱卵量10粒左右,平均产卵量10~15粒左右;食性杂,群居危害,喜食白茎盐生草(Halogeton arachnoideus)等沙生草本植物的种子。【结论】该虫具有以成虫形态越冬和滞育、迁移性强等生物学特性。在生态环境发生较大变动后,能够经历逆境后迅速恢复种群并迁移危害。宁夏中南部地区种植结构调整,极大改变了该地区的荒漠生态环境,促使巨膜长蝽在经过夏季高温滞育恢复种群后无法获得足够野生寄主的情况下,大规模迁入农田系统危害。