Simultaneous Expression of <Emphasis Type="Italic">Spinacia oleracea</Emphasis> Chloroplast Choline Monooxygenase (CMO) and Betaine Aldehyde Dehydrogenase (BADH) Genes Contribute to Dwarfism in Transgenic <Emphasis Type="Italic">Lolium perenne</Emphasis>

Choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH) catalyze the first and second steps in the biosynthesis of glycine betaine in betaine-accumulating plants. Over-expression of the Spinacia oleracea chloroplast choline monooxygenase (SoCMO) and betaine aldehyde dehydrogenase (SoBADH) genes has not been reported in Lolium perenne. In this investigation, the SoCMO and SoBADH genes have been used to generate transgenic L. perenne plants via particle bombardment. Transgenic plants have been confirmed with PCR, Southern blot, and Northern blot analyses. Enhanced salt stress tolerance has been observed from SoBADH–SoCMO transgenic L. perenne plants. The dwarf phenotype was first observed 3 months after transgenic plants were established in soil and was to be stably inherited. Height of transgenic plants was decreased by 63% compared to the control. Measurement of endogenous GAs content demonstrated that the content of endogenous GA1 was decreased by 75.2%, and the content of endogenous GA4, GA12, GA19, and GA53 of transgenic plants was increased by 200%, 221%, 105%, and 108%, respectively, compared to the control plants. Dwarf trait of SoBADH–SoCMO transgenic L. perenne plants can be recovered by application of exogenous GAs. These results demonstrated that simultaneous expression of the SoCMO and SoBADH genes enhanced salt stress tolerance and induced dwarfism in transgenic L. perenne. Dwarfism induced by expression of the SoCMO and SoBADH genes was associated with synthesis of endogenous GAs and it could be recovered by application of exogenous GAs. This is the first report on dwarfism induced by expression of the SoCMO and SoBADH genes in a species in turfgrass.     

An efficient callus suspension culture system for triploid bermudagrass (Cynodon transvaalensis × C. dactylon) and somaclonal variations

An efficient callus suspension culture and regeneration system in a triploid bermudagrass (Cynodon dactylon × C. transvaalensis cv. Tifeagle) was studied in this report. Proline improved callus proliferation, but had no effect on regeneration. 0.6–1.2 mg l⁻¹ BA improved regeneration, but higher concentrations of BA (≤1.2 mg l⁻¹) resulted in the production of rootless plantlets. The embryogenic calli were able to proliferate continuously for at least 2 years with regeneration ability through the established suspension culture system. Observations with scanning electron microscope and light microscope showed somatic embryogenesis during the regeneration. Somaclonal variations were observed in regenerated plants. More than 2000 regenerants were screened for drought tolerance in the greenhouse, from which seven lines appeared to have increased drought tolerance relative to their parental variety. It is suggested that somaclonal variation in triploid bermudagrass offers an effective tool for its breeding.     

CBF/DREB转录因子与植物矮化的相关性研究进展

CBF/DREB转录因子即干旱应答元件结合蛋白,是一类可以调控多个与干旱、高盐及低温耐性有关的功能基因表达的转录因子家族。很多报道称CBF/DREB转录因子的过量表达使转基因植株产生矮化、晚花现象。着重探讨CBF/DREB转录因子与植物矮化现象相关性与其矮化机理,并对草坪草育种新方向进行展望。     

Effects of Cytokinin and Nitrogen on Drought Tolerance of Creeping Bentgrass

Cytokinin (CK) is a vital plant hormone that controls many aspects of growth and development in plants. Nitrogen (N) is the indispensable macronutrient needed in plants and also one of the most important limiting factors for plant growth. This study was designed to investigate the simultaneous effects of CK and N on the visual turf quality and antioxidant metabolism of drought-stressed creeping bentgrass (Agrostis stolonifera L.). ‘PennA-4’ creeping bentgrass treated with trans-zeatin riboside at three rates of CK concentrations of 0, 10 and 100 μM (designated by CK0, 10, and 100) and two nitrogen rates with 2.5 and 7.5 kg N·ha^-1 every 15 days (designated by low and high N) in a complete factorial arrangement was grown under two soil moisture regimes: well-watered and drought stress. Exogenous CK improved turf quality and delayed leaf wilting under drought stress, especially under high N. The grasses treated with CK10 and CK100 had lower O₂^- production and H₂O₂ concentration than those without CK treatment. The CK100 treatment enhanced the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and guaiacol peroxidase (POD) by 25%, 22%, 17% and 24%, respectively, relative to CK0. Moreover, the activity changes of the antioxidant enzyme isoforms were more significant under high N condition relative to low N condition. Our results demonstrated the beneficial impacts of CK and N on physiological reactions, especially antioxidant metabolism, and foliar application of CK at 10 or 100 μM plus 7.5 kg ha^-1 N biweekly may improve drought stress resistance of creeping bentgrass.     

植物细胞悬浮培养影响因子研究

植物细胞悬浮培养已广泛应用于育种和生产次生代谢产物,在生物工程中发挥着重要作用。本文综述了愈伤组织继代改良的措施,影响胚性悬浮细胞系建立的几个因素,存在问题及前景展望。     

一种高质量麦冬总RNA的提取方法

本方法采用CTAB作为去污剂,分别用氯仿/异戊醇反复抽提、LiCl沉淀,以去除蛋白质、碳水化合物和次生代谢物等杂质,用DNase处理去除DNA污染,最后用无水乙醇沉淀获得总RNA。该方法不仅能获得完整性好、纯度高的总RNA,而且操作简单、成本低廉、RNA产率高,对富含次生物质的中草药材植物组织总RNA的提取具有借鉴意义。     

麦冬基因组DNA提取方法研究

为了从富含多糖、多酚的顽拗植物麦冬中分离出高质量基因组DNA,分别建立了改良CTAB法和改良SDS法。通过使用核分离液和CTAB/NaCl溶液、提取液中加入0.35倍体积无水乙醇等最大限度地除去杂质。将两种方法提取的8种麦冬DNA与两种离心柱式试剂盒提取的DNA在纯度、产率等方面进行对比,并进行双酶切和SRAP分析。麦冬类植物SRAP反应体系的建立尚属首次。结果表明,改良CTAB法和改良SDS法均能从8种麦冬叶片中提取到高质量的基因组DNA,改良CTAB法提取纯度更高,提取幼叶DNA纯度可以与离心柱式试剂盒媲美,能够满足多种分子生物学试验要求。在后续试验中,用改良CTAB法从20多种沿阶草族植物中提取到了高纯度DNA。该方法通用性好,提取的DNA纯度高,对其他顽拗类植物基因组DNA的提取具有借鉴意义。     

Study on optimization of moisture retention for golf green rootzone soil mixtures

Most golf course green have been constructed with pure sand or sand-based rootzone mixes. As we know, high sand content provides rapid drainage despite sand’s inefficiency in retaining moisture. However, drainage capability and water retention are both essential elements to the golf course green, and the addition of peat could increase the soil moisture retention, therefore, the research on the drainage capability and water retention of the sandy golf green has become more and more important these years. In this study, extreme vertex design was applied which is one of the mixture experiment designs widely used in mixture experiments, the study investigated the effects of the thirteen different rootzone soil mixtures using middle-coarse, fine sand, very fine sand plus silt and clay as well as peat as the materials under three kinds of golf green profile (1-layer profile, 2-layer profile, 3-layer profile) conditions on the water retention of green rootzone. Through the qualitative, quantitative and optimization analysis of water retention capability of the sandy golf green, evidence a basis for choice of green profiles and rootzone matrix could be provided. And the significantly correlative regression model was established between the moisture retention and components of rootzone soil mixture. In addition, the order of factor contribution ratio, effect of single and double factor and optimization of the model were analyzed in detail. The results were as follows: both green profile and soil mixture, which had interaction of each other, had significant effects on soil moisture retention. Additional attributes include high porosity and greater water holding capacity than sand, and the higher content of peat, fine sand plus silt and clay, the better water retention. The mixtures had much higher water content in 1-layer profile than that in the other two profiles. There was significantly higher water content in 2-layer profile for pure sand mixtures (A–E) and low peat mixture (F) than that in 3-layer profile, while there was no higher water content for other 7 high peat mixtures (G–M) (>5%) in 2-layer than that in 3-layer profiles. The significance of key factors in rootzone soil mixture on moisture retention were: very fine sand plus silt and clay > peat > middle-coarse > fine sand. According to the moisture retention 15–25% specification of USGA (United States Golf Association), the optimal soil mixture in 1-layer profile was: middle-coarse 71.4–73.5%; fine sand 17.8–21.5%; very fine sand plus silt and clay 6.8–8.4%; peat 0–1%. The optimal soil mixture in 2-layer profile was: middle-coarse 65.0–73.4%; fine sand 17.8–20.5%; very fine sand plus silt and clay 7.5–8.9%; peat 0.2–6.3%. The optimal soil mixture in 3-layer profile was: middle-coarse 62.3– 73.9%; fine sand 17.7–21.4%; very fine sand plus silt and clay 7.3–10.7%; peat 0–6.3%. These optimal recipes took through the limitation of previous research, which were practically important to golf green soil selection and profile design. Thus, both proportion and interaction should be considered when we choose the soil mixture.     

植物DREB转录因子与植物激素的相互作用

DREB是植物中重要的转录因子,调控一系列非生物胁迫相关基因的表达,增强植物抵抗环境胁迫的能力.同时,当植物受到逆境胁迫时,植物体内的酶和激素都会发生变化,从而影响一系列的生理活动或生化变化来产生抵御胁迫的适应能力.近些年来,研究者们发现DREB与植物激素之间关系密切,从植物激素角度入手研究DREB的功能逐渐成为热点.就逆境胁迫下DREB转录因子与激素之间的相互作用进行阐述.