甘蔗茎尖脱毒培养技术研究

针对热处理结合茎尖分生组织培养甘蔗脱毒苗在茎尖培养阶段由于污染率和褐变率高培养成功率较低问题,在甘蔗的热处理、茎尖解剖、接种等阶段设计不同试验进行研究探讨,以期提高茎尖培养的成活率和成功率。试验结果表明,不同的甘蔗品种在热处理时应需不同的水浴温度和时间;在茎尖解剖前,使用75％酒精处理可以减少茎尖培养的污染率;在接种前,将茎尖放到无菌水或者150mg／L的PVP（聚乙烯毗咯烷酮）溶液中浸泡15min可以减少酚污染,PVP还对其生长有促进作用;活性炭的作用有利有弊,需要进一步的研究确定其用量。     

花生种间杂种胚胎发育及内源激素变化

以花生栽培品种豫花15为母本分别与野生种A. correntina和A. macedoi杂交,以栽培品种间杂交组合豫花15×白沙1016为对照,采用石蜡切片技术观察花生杂种胚胎发育进程,酶联免疫吸附分析法(ELISA)测定花生杂种胚胎内源激素含量, 分析胚胎发育进程与胚胎内源激素含量的关系。结果显示,3个杂交组合的下针率无明显差异,表明栽培种与2个野生种杂交不存在受精前障碍;与对照相比,豫花15×A. correntina表现杂交可亲和,其胚胎发育中激素含量的变化与对照组合基本一致;而豫花15×A. macedoi杂交授粉后12 d内胚胎发育基本正常,17~27 d发育缓慢,之后珠被迅速向内增生抑制了胚的进一步发育并最终败育,在同一时期的杂种胚胎中,3-吲哚乙酸(IAA)、赤霉素(GA₃)和玉米素核苷+二氢玉米素核苷(ZR+DHZR)等促进生长的激素维持在较低的水平,而脱落酸(ABA)含量以及脱落酸与某一促进生长激素或3类促进生长激素之和的比值保持了较高的水平,推测可能是激素代谢的失衡导致种间杂种胚胎的败育。用IAA、激动素(KT)单独或混合处理不亲和组合豫花15×A. macedoi杂交授粉后的果针基部表明,外施促进生长的激素能够明显增加胚珠的长度,一定程度上验证了上述推测。     

轮缘间隙对轴流泵内部非定常流场的影响

为研究轴流泵轮缘间隙泄漏流的非定常特征及其对泵外特性的影响,采用基于S-A模型的DES方法和滑移网格技术,对轴流泵在设计流量下的内部湍流进行了数值计算,重点分析了4组轮缘间隙下泵内非定常流场特性及压力脉动特性.在设计轮缘间隙下,计算所得泵扬程和效率与试验数据吻合良好,最大相对误差分别为2.0%和3.0%.计算结果表明:随轮缘间隙增大,水泵扬程和效率均呈下降趋势;轮缘泄漏涡强度和影响范围随轮缘间隙增大而增大,当轮缘间隙为3.3‰D₂时,轮缘泄漏涡扩散至相邻叶片出口边;不同轮缘间隙下,叶轮区压力脉动频率均以叶频为主;靠近叶片进口的叶轮室内壁压力脉动幅值随轮缘间隙的增大呈减小趋势,叶轮室中部压力脉动随轮缘间隙增大而增大;叶轮出口断面的压力脉动频域特性在不同轮缘间隙下均以1倍叶频为主,脉动幅值随轮缘间隙增大而减小.     

改良大豆胚尖再生体系的研究

为了研究利用Thidiazuron(TDZ)建立大豆胚尖高效再生体系的可行性。跃进10号的成熟种子浸泡16 h后,收集胚尖转入添加了不同浓度的TDZ芽诱导培养基中,4周后计算出芽率,并对拔高培养基和生根培养基进行优化。TDZ浓度为0.08 mg/L时,出芽率最高,达92.6%,平均每个外植体产生的丛生芽的数目也最多。适宜大豆胚尖的芽诱导分化培养基为MS+0.08 mg/L TDZ,拔高培养基为MS+2.0 mg/L KT+0.2 mg/L NNN,生根培养基为1/2MS+1.0 mg/L IBA。利用TDZ建立大豆胚尖高效再生体系是可行的,提高了丛生芽的分化率,为快繁和提高遗传转化效率奠定了坚实的基础。     

Construction of mouse ESC line stably expressing Pdx1 by Tet-On system

AIM To construct the mouse embryonic stem cell （ESC） line with stable pancreatic and duodenal homeobox 1 （Pdx1） expression by Tet-On system, which may lay a foundation for further research on the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells. METHODS The Pdx1-overexpressing lentiviral vector with green fluorescent protein marker and puromycin resistance was constructed by Tet-On system and was used to infect the mouse ESC. The cells were divided into 3 groups： blank control group （ESC group）, empty lentivirus control group （PDX1^- ESC group） and Pdx1 lentivirus transfection group （PDX1⁺ ESC group）. Flow cytometry was used to detect the transfected cells after screening by doxycycline （DOX）. The function of Tet-On system and the expression of Pdx1 gene were detected. The transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were sorted by flow cytometry, and constructed ESC line with stable expression of Pdx1 and negative control ESC line were verified. RESULTS （1） The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were 90.72% and 94.01% after screening by DOX, respectively. The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group was 97.84% and 98.13% after sorting by flow cytometry, respectively. （2） With DOX, green fluorescence was observed in PDX1^- ESC group and PDX1⁺ ESC group. The mRNA and protein expression of Pdx1 was significantly increased in PDX1⁺ ESC group （P<0.05）. Without DOX, no green fluorescence was observed in the cells of the 3 groups, and no significant difference in the mRNA and protein expression of Pdx1 was observed （P>0.05）. （3） After 3 months of cryopreservation, the cell lines still survived in resuscitation culture and were regulated by DOX. CONCLUSION Using Tet-On system, the mouse ESC line with inducible Pdx1 expression were successfully established and could be used as an effective cell model to research the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells.     

Differential Sensitivity of Rice Cultivars to Salinity and Its Relation to Ion Accumulation and Root Tip Structure

Abstract

Effects of NaCl on the growth, ion content, root cap structure and Casparian band development were examined in four rice (Oryza sativa L.) cultivars with different salt resistance (salt-sensitive indica-type IR 24 and japonica-type Nipponbare and salt-resistant indica-type Nona Bokra and Pokkali). Experiments were conducted to find the differences in salinity resistance during early seedling and developed seedling stages among the cultivars. For salinity treatment, sodium chloride (NaCl) was added to nutrient solution at concentrations of 0, 25 and 50 mM for 7 days from germination to the 7th day (early seedling stage) or from the 7th day to 14th day (developed seedling stage). Growth inhibition by salinity was more prominent in the early seedling stage than in the developed seedling stage. Based on the growth, the order of the sensitivity was IR24 > Nipponbare > Nona Bokra > Pokkali. The growth of NaCl-treated rice cultivars relative to control was significantly and negatively correlated with the Na⁺ content and Na⁺/K⁺ ratio in roots and shoots in both stages. Scanning electron microscopic observation revealed that the root cap tissues proliferated and extended to the basal part of the root tip by salinity. The length of root cap was, however, reduced by 50 mM NaCl in sensitive cultivars due to peeling off. An endodermal Casparian band was formed in the basal region of the root tip. Development of the Casparian band was more prominent in sensitive cultivars than in tolerant cultivars. Root cap proliferation might be related to NaCl resistance in rice seedlings, but the Casparian band may not function efficiently in Na⁺ exclusion. Essentially the present results suggest that exclusion of Na⁺ from roots plays a critical role in expression of Na⁺ resistance in rice seedlings and the root cap is important for Na⁺ exclusion.     

利用近等基因系研究穗分枝小麦的生长锥发育及其物候特征

利用一对穗分枝差异的小麦近等基因系,研究其生长锥分化过程和发育规律。结果表明:穗分枝小麦发育至二棱期后,分枝穗轴突起形成,继而形成穗分枝;发育过程可分为9个时期:初生期、伸长期、单棱期、二棱期、分枝突起形成期、分枝小穗突起形成期、小花分化期、雌雄蕊分化期和抽穗始期;物候学方面,分枝小麦在二棱期之前的发育进程与正常穗型小麦一致,但分枝穗轴形成后,分枝小麦的拔节期、抽穗期和开花期分别要晚于正常穗型4.2、2.3 d和1.4 d,说明穗分枝小麦二棱期之后有一个分枝穗轴突起形成和伸长期。本研究有利于了解穗分枝小麦生长锥的发育进程,为进一步实现小麦遗传改良、优化栽培技术和加快该种质资源的利用奠定形态学基础。     

甜高粱组织再生研究

通过对不同外植体的筛选和培养条件的优化,建立了甜高粱再生体系。结果表明,以茎尖和茎为外植体的培养可获得甜高粱再生苗,但分化率较低,褐化较为严重;而以叶和根为外植体的培养不能获得甜高粱再生苗;成熟胚作外植体分化率高达98.7%,且未发生褐化。     

灾害蝗虫附着系统爪尖的剪切强度测算

蝗虫爪尖能够形成用于稳固附着的机械锁合,抑制蝗虫在捕集滑板表面的滑移行为。为获取用于蝗虫捕集滑板研制的爪尖强度信息,利用自行设计的昆虫微力测试系统测试了蝗虫前足、中足、后足爪尖的剪切力;采用扫描电镜(scanning electron microscopy,SEM)分析了蝗虫爪尖及其断裂面的微形貌结构与几何特征,计算了爪尖断裂面的截面积,并据此测算了蝗虫爪尖的剪切强度。结果表明爪尖的剪切力介于197.4~243.6 mN,其中前足爪尖的剪切力最大,后足居中,中足最小;爪尖断裂面表现出层状的复合结构且具有显著不同的截面积(10 847.6~13 908.5μm2);测算结果表明蝗虫爪尖的剪切强度介于14.2~22.5 MPa。该研究可为研制具有良好滑移功能蝗虫捕集滑板提供参考,并可促进灾害蝗虫光电诱导滑移捕集技术的发展。     

文心兰茎尖离体培养研究

文心兰茎尖培养的研究，试验结果表明：文心兰茎尖在MS BA2mg／L NAA0．5mg／L培养基上，离体培养1个月后产生原球茎．改良KC基本培养较适原球茎的增殖；用BAI．5mg／L NAA0．2mg／L的激素配比，原球茎月增殖宰可达332．5％；CW可以减少原球茎褐死现象和促进原球茎的增殖：壮苗阶段，以附加NAA0．5% 10％香蕉汁的效果较好．水苔是良好的移栽基质，成活率在85．3％．