外源DNA导入对小麦转基因D1代种子萌发和植株生长的影响

通过花粉管通道介导玉米、稗草总DNA导入4个冬小麦品种(系)9411,955159,96266,鲁麦21中,研究外源DNA导入对转基因D1代种子萌发和植株生长的影响.结果表明:各受体的转基因D1代T1(导入玉米DNA)、T2(导入稗草DNA)处理的种子发芽势和发芽率与对照ck相比呈显著或极显著降低,而T1和T2间无显著差异;以冬小麦9411和鲁麦21为受体的T1株高和ck相比差异极显著,而T2株高与ck间差异不显著;96266为受体的T1处理的第1节间长与对照相比差异显著;9411 T1和T2处理稳长与ck相比差异均达到极显著水平.     

外源C_4作物DNA导入小麦的研究

利用小麦自交后的花粉管通道,将C_4作物玉米或高粱的DNA导入小麦的种胚细胞,结果受体后代发生了广泛的变异,而且受体不同,所发生的变异不同。甘麦8号受体后代植株变矮,结实率低,分蘖多至5个,少至完全不分蘖;8347—2-4和8013—3品系后代中出现不育株;6873—20—3品系受体后代的籽粒由红色转变为白色而且稳定遗传;蓝粒麦受体后     

野生稻DNA导入水稻栽培品种获得变异种质的农艺性状及抗病性初步分析

利用花粉管通道导入技术将普通野生稻（从染色体组）DNA导入宁夏水稻栽培品种宁粳23号、宁粳16号中，获得163个变异后代，变异性状极其广泛，对其中22份材料在全生育期经苗期、成株期抗病性鉴定，表现广谱抗性的材料有4份，其中2005D3-136、2005D3-60无论在农艺性状还是在抗病性上变异都优于受体。     

水稻高世代回交导入系耐盐性的遗传研究

水稻是一种对盐敏感的作物，盐分是限制水稻产量的主要因子之一。本研究利用广泛推广种植的杂交籼稻的亲本珍汕97（ZS97）和测序粳稻品种日本晴（Nipponbare）杂交、回交构建的导入系，对水稻的耐盐性进行遗传研究。用88份导入系作苗期盐胁迫（0．3％氯化钠）处理试验，发现6份导入系的耐盐性同受体亲本ZS97有显著差异，其中5份导入系耐盐，1份对盐敏感。图示基因型分析表明5份导入系中含有少数外源导入片段及耐盐相关的基因。     

野生稻DNA导入栽培品种获得变异后代的抗病性分析

利用花粉管通道导入技术将普通野生稻(AA染色体组)DNA导入宁夏水稻栽培品种宁粳23号、宁粳16号中,获得163个变异后代,变异性状极其广泛,对其中22份材料在全生育期经苗期、成株期抗病性鉴定,表现广谱抗性材料有4份,其中2005D3-136,2005D3-60无论在农艺性状还是在抗病性上变异都优于受体。     

玉米DNA导入水稻变异后代的研究

利用花粉管通道法将以玉米DNA导入水稻品种龙粳4中,以提高水稻的光合效率。对生育中、后期叶绿素a、b和叶绿素含量、净光合速率、DNA差异、过氧化物酶及脂酶同工酶电泳和农艺性状的研究表明：与受体相比,外源DNA导入引起变异后代在生理、DNA、蛋白质和形态性状上发生改变,从而引起光合特性表现不同程度的变异。证明通过导入可以将玉米的高光效特性传递到水稻中,为常规育种与生物技术相结合选育高光效水稻新品种提供依据。     

大麦DNA导入小麦诱导抗白粉病变异的研究

本试验利用外源DNA导入技术,研究了大麦DNA导入小麦品种后,D_1,D_2和D_3代的抗白粉病变异株(系)在不同条件下的抗性表现及产量构成、籽粒蛋白质和氨基酸含量的变化。试验结果表明,导入外源DNA的D_1代小麦檀株出。现了抗白粉病等多种变异类型,其中免疫和高抗白粉病变异株古2.77％,且抗性能够向子代传递,其D_2代在大田自然发病和温室接菌条件下,有5个株系抗性保持稳定,8个株系有分离,其中一个株系在D_2和D_3代抗性均稳定。在田间D_2代有2个稳定株系(D_2-20,D_2-29)的籽粒粗蛋白质含量,比受体分别高20.3％和15.76％,17种氨基酸总量分别高23.4％和27.5％。在温室这些性,状的数值也明显高于受体。     

λDNA导入中国春后花粉母细胞减数分裂染色体行为及性状变异观察

将λDNA导入中国春后,在当代花粉母细胞（PMC）减数分裂过程中,观察到了单价体、染色体落后、染色体桥、微核、细胞多极分裂等异常现象,发生异常现象的PMC约占观察占PMC的25．54％。这些染色体异常可能是异源DNA整合到受体染色体,进行染色体重排所引起的复杂细胞学反应。后代植株在茎秆硬度、株高、穗长、穗形、小穗数、穗粒数、麦芒有无、结实率、成熟期等形态性状方面发生了明显的变异。发生变异的可能机制是异源遗传物质进行细胞核以后,通过重排整合到了受体的染色体上,从而激活受体本身的一些沉默基因,或者抑制了一些基因的表达;导入的异源基因直接表达,改变了受体后代的表现型;异源DNA导入受体后引起了染色体结构变异。     

玉米衔接式单片段导入系群体的构建和评价

以生产上广泛种植的玉米杂交种豫玉22的亲本自交系87-1和综3为受体亲本,以糯质、抗病性较好的玉米自交系衡白522为供体亲本,采用回交和自交的方法,结合SSR分子标记辅助选择,分别构建了以87-1和综3为背景的衔接式玉米单片段导入系群体,并对其遗传背景、导入片段大小、数目和覆盖率等进行了评价。结果表明,87-1背景的导入系群体导入了40个供体片段,片段长度介于0.03~342.8 cM,平均长度为91.1 cM,导入片段总长为3 643.9 cM,覆盖率为48.9%;综3背景的导入系群体导入了78个供体片段,片段长度介于0.03~343.4 cM,平均长度为75.5 cM,导入片段总长为5 895.2 cM,覆盖率为79.2%。     

外源DNA导入水稻后代变异性的SSR分析

采用花粉管通道法,将高粱DNA导入水稻,获得了34个稳定的变异品系,从60对SSR引物中筛选出17对,对供体高粱、受体水稻及其外源DNA导入后代稳定材料进行了SSR分析,结果表明,高粱DNA导入水稻可以引起广泛的变异,在分子水平上产生了遗传多样性,聚类分析可将其分为五类,各类有其特有的遗传变异性。接受外源DNA三个片断,两个片断及一个片断的株系分别被聚成一类,与其他类的遗传差异分别为0.467,0.389及0.347,变异程度依次递减;未接受外源DNA两个片断的株系被聚成另一类。因此,接受外源DNA片断的多寡是水稻后代稳定品系变异程度的基础,也是它们分类的标准。     

2种种植方式下不同化感杂交稻组合和新品系对稗草的抑制作用

以美国强化感稻‘PI312777’为对照,比较不同化感杂交稻组合和新品系抑制稗草能力的田间表现,探索利用水稻化感作用的有效方法。采用移栽和直播2种水稻种植方式,对水稻封行前抑草率、竞争抑草率、灌浆期抑草率、伴生稗草平均株高和稗草单株干物重进行调查分析。化感稻移栽条件下对于稗草株高和干物重的抑制效果优于直播条件;直播条件下化感稻的化感抑草率大于移栽条件下,且变异系数较小,可见直播方式更利于化感稻种植;环境条件是化感稻抑草作用发挥的重要影响因子,其中竞争抑草率受环境的影响较大,化感作用表现相对稳定;相关分析表明化感稻的抑草作用并未对存活稗草的地上部分生长量有明显影响,增强化感稻的化感作用,能有效抑制田间稗草的萌发。实验结果显示了化感稻是更适于直播种植的材料,在生产上具有良好的应用前景。     

从化感水稻根际土壤中筛选抑草细菌的研究

从化感水稻PI312777根际土壤中筛选具有抑草作用的细菌,为开发抑草微生物资源提供科学依据。以稀释涂布平板法筛选菌株,以莴苣、稗草为受体,实验室生物测试和盆栽试验相结合,确定候选菌株的抑草活性。实验室生物测试结果表明,有2个菌株Z1和Z2的发酵液对莴苣具有强抑制作用,且对稗草根长的抑制率为50%~60%,株高的抑制率为40%~50%。盆栽试验结果表明,Z1和Z2的发酵液对稗草根长的抑制率分别为28.1%和27.0%,株高的抑制率分别为8.85%和4.48%。菌株发酵液对3种水稻的根长有促进作用,对株高没有影响或小的抑制作用（抑制率3%）。结果表明水稻化感抑草作用与根际微生物有关,暗示从化感水稻根际土壤中筛选抑草细菌是一个可行的思路。     

水氮互作下长雄野生稻化感作用与田间抑草效果

以化感抗稗草长雄野生稻(Oryza longistaminata),非化感栽培稻RD23以及它们的F₁代(RD23 × O. longistaminata)为材料,采用实验室内生物测定和田间调查相结合方法,研究水肥对长雄野生稻化感作用的影响和探讨对野生稻化感作用及其田间抑制杂草作用的互作关系。在移栽后20~50 d内设淹水、2种形式的干湿交替和旱种等4种田间管理方式,对每种水分管理方式分别设3个施氮(尿素)水平处理。野生稻叶片的水提液,检测了对稗草的化感作用。结果表明,长雄野生稻化感作用在干旱与不施氮水平下最强,对稗草根长与干重的抑制率分别达到69.3%和74.6%,但随着施氮水平的提高与淹水时间的延长而降低;田间则以干湿交替条件下控制稗草效果最好,旱种管理后进行灌水能显著提高野生稻控制稗草的效果。水分与氮互作效应对长雄野生稻化感作用及其田间抑制杂草效果极显著,对F1代化感作用及其田间抑制杂草效果也达显著水平。     

恢复系752近等基因导入系构建与初步利用研究

利用国际水稻所提供的127份核心种质资源作为供体亲本,与我所育成的籼型优良恢复系752为受体亲本作杂交与回交,构建了3300份近等基因导入系。初步研究结果表明,在近等基因导入系中可以筛选到许多有利基因如抗稻瘟病、耐低氮、耐低磷和耐旱性等;且其中95％的导入系对野败籼型不育系具有良好的恢复能力,表现出较强的杂种优势。因此,利用核心种质资源改良当地的优良亲本,可以培育出符合生产目标的水稻优良新品种。     

非洲栽培稻基因渗入系的稻米品质鉴定与筛选

稻米品质改良越来越成为现代水稻育种工作的重要目标,探明新选育非洲栽培稻基因渗入系的稻米品质潜力将为其在水稻优质育种工作中的进一步应用奠定基础。对50份非洲栽培稻基因渗入系的12个稻米品质性状进行了考察鉴定,并结合相关性分析、聚类分析和判别分析方法对其综合品质表现进行了分类评价。供试材料均表现较高的糙米率(≥74.19%)、精米率(≥67.83%)、较大的长宽比(≥2.79)和低水分含量(≤13.03%),多数选系具有适宜的直链淀粉含量(17.00-22.00%)。聚类分析将供试材料分为5组,其中第I组的综合米质性状最优;典型判别分析获得2个典型判别函数,分别可解释89.5%和7.2%的方差变异,主要反映垩白粒率和长宽比等指标。新选育的非洲栽培稻基因渗入系在糙米率、精米率、长宽比和直链淀粉含量方面具有优势,垩白粒率和长宽比是非洲栽培稻基因渗入系品质改良的重要指标；本研究将为非洲栽培稻基因渗入系在高产优质水稻新品种选育中的研究应用提供依据。     

稻田土壤中抑草真菌的筛选与鉴定

以稻田土壤为材料,采用稀释涂布法,结合生物抑草活性测试跟踪,筛选分离抑草真菌微生物。结果表明：从稻田土壤中筛选到32个菌株,其中11个菌株对莴苣根长和株高的抑制率高于50%,这11个菌株中有5个对稗草株高的抑制率高于40%,而对水稻具有抑制作用的有3株,没有抑制作用的2株。8号菌株因对稗草具有较好抑制率而对水稻没有影响,作为目标菌株,开展了其生长特性研究。形态和分子生物学鉴定表明8号菌株属于曲霉属。该菌株的最优培养条件为以蔗糖为碳源,以酵母浸膏或蛋白胨为氮源,最适温度30℃,最适pH 6。8号菌株的菌液含蛋白质5.14 mg/L,总酚酸5.59 mg/L,可溶性糖448.12 mg/L,总黄酮为2.39 mg/L。8号菌株可开发利用为微生物除草剂应用于稻田除草。     

稗草质源水稻渝819B光合特性分析

稗草是水稻属间植物，携带有水稻需要的有利基因。为了研究稗草外源基因导入后对水稻光合特性的影响，利用花粉管通道技术将稗草DNA导入到水稻保持系‘渝805B’中选育出水稻新保持系‘渝819B’，通过对‘渝819B’和背景亲本‘渝805B’在功能叶SPAD值和光合指标、产量结构及其组合产量进行比较分析，结果表明，‘渝819B’在叶绿素含量、净光合速率、胞间CO2浓度、产量结构及所配组合的产量均明显优于‘渝805B’，而在光合指标的在蒸腾速率、气孔导度性状上又低于‘渝805B’的相应值。说明稗草质源‘渝819B’具有典型的高光效特性。     

Identification and molecular cytology analysis of cold tolerance introgression lines derived from <Emphasis Type="Italic">Oryza sativa</Emphasis> L. mating with <Emphasis Type="Italic">O. rufipogon</Emphasis> Griff.

A previously constructed introgression lines (ILs) population including 239 lines was used to evaluate the cold-tolerant ability. The ILs with strong cold tolerance which were identified in the present study were used for further cytological and microsatellite (SSR) marker analyses. The results showed that the IL5243 and IL5335 had strong cold-tolerant ability. Cytological analysis showed that the rate of pollen mother cells (PMCs) with normal meiotic behavior in the IL5243 and IL5335 was to 89.93 and 90.22 %, respectively, and finally formed normal mature pollen. And meanwhile, the low frequency of abnormal chromosome behavior was observed in the IL5243 and IL5335, such as univalent, 8-shape bivalent, multivalent at diakinesis. At anaphase I, one or two lagging chromosomes were observed in some PMCs (3.95–5.15 %). The results of SSR marker analysis further confirmed that partial alien DNA of common wild rice has been transferred into the IL5243 and IL5335. These results implied that the IL5243 and IL5335 might be excellent bridging germplasm for exploring and utilizing the cold-tolerant gene of common wild rice. In addition, IL5243 and IL5335 would provide a better experimental system for understanding some epigenetic phenomenon induced by alien gene introgression.     

Herbicidal Potential and Quantification of Suspected Allelochemicals from Four Grass Crop Extracts

Unknown compounds in crop plants are inhibitory to seed germination and early seedling growth of weed plants. A Petri dish assay showed that barley (Hordeum vulgare L.), oats (Avena sativa L.), rice (Oryza sativa L.) and wheat (Triticum aestivum L.) extracts significantly reduced root growth of alfalfa (Medicago sativa L.), barnyard grass (Echinochloa crus‐galli, Beauv. var. oryzicola Ohwi.) and eclipta (Eclipta prostrata L.). As the concentration of crop extracts increased, root growth of the test plants were significantly reduced. A high‐performance liquid chromatography analysis with nine standard phenolic compounds showed that the concentrations and compositions of allelopathic compounds depend on the extracted plant extracts. Caffeic acid, hydro‐cinnamic acid, ferulic acid, m‐coumaric acid, p‐coumaric acid and coumarin were present in all the crop plant species, and hydro‐cinnamic acid were detected as the highest amount. Coumarin at 10^?3 m significantly inhibited root growth of alfalfa and barnyard grass more than that of eclipta. The research suggests that extracts of barley, oats, rice and wheat have an allelopathic effect on alfalfa, barnyard grass and eclipta and that the findings of bioassay were considerably correlated with the type and amount of causative allelochemicals, indicating that the allelopathic effects on three test plants were ranked in order of wheat (highest), barley, rice and oats (lowest). The results may have value in enabling weed control based on natural plant extracts or crop residues in the fields.     

Development of high yielding IR64 × <Emphasis Type="Italic">Oryza rufipogon</Emphasis> (Griff.) introgression lines and identification of introgressed alien chromosome segments using SSR markers

Modern rice varieties that ushered in the green revolution brought about dramatic increase in rice production worldwide but at the cost of genetic diversity at the farmers’ fields. The wild species germplasm can be used for broadening the genetic base and improving productivity. Mining of alleles at productivity QTL from related wild species under simultaneous backcrossing and evaluation, accompanied by molecular marker analysis has emerged as an effective plant breeding strategy for utilization of wild species germplasm. In the present study, a limited backcross strategy was used to introgress QTL associated with yield and yield components from Oryza rufipogon (acc. IRGC 105491) to cultivated rice, O. sativa cv IR64. A set of 12 BC₂F₆ progenies, selected from among more than 100 BC₂F₅ progenies were evaluated for yield and yield components. For plant height, days to 50% flowering and tillers/plant, the introgression lines did not show any significant change compared to the recurrent parent IR64. For yield, 9 of the 12 introgression lines showed significantly higher yield (19–38%) than the recurrent parent IR64. Four of these lines originating from a common lineage showed higher yield due to increase in grain weight and another three also from a common lineage showed yield increase due to increase in grain number per panicle. For analyzing the introgression at molecular level all the 12 lines were analyzed for 259 polymorphic SSR markers. Of the total 259 SSR markers analyzed, only 18 (7.0%) showed introgression from O. rufipogon for chromosomes 1, 2, 3, 5, 6 and 11. Graphical genotypes have been prepared for each line and association between the introgression regions and the traits that increased yield is reported. Based on marker trait association it appears that some of the QTL are stable across the environments and genetic backgrounds and can be exploited universally.