Cu^2＋浓度对啤酒大麦幼胚组织培养与植株再生的影响

为了提高大麦幼胚胚性愈伤组织和植株再生频率,本试验以引进的4个啤酒大麦品种为材料,在诱导、分化与生根培养基上分别进行了不同浓度Cu2+处理.结果表明,MS诱导培养基中增加Cu2+浓度对啤酒大麦愈伤组织的诱导无明显影响,但对胚性愈伤组织的形成有明显的促进作用;在分化与生根培养基中不同浓度Cu2+处理的各品种再生率也存在显著差异,都表现为在5.0～10.0μmol@L-1的浓度下明显提高了出愈率和再生率,而且每个胚性愈伤组织的平均再生率也有显著提高.通过切片技术证明Cu2+浓度主要影响胚性愈伤组织的发生.供试品种在出愈率和再生率上也存在一定的差异,同时Cu2+浓度和基因型之间表现出正互作效应.     

大豆不同外植体植株再生的研究

采用不同大豆品种的下胚轴、上胚轴、小真叶和幼胚作外植体进行植株再生的研究。结果表明汾豆３３号诱导植株再生率最高;诱导频率依次为下胚轴＞上胚轴＞小真叶＞幼胚;下胚轴在Ｂ５＋ＮＡＡ０．３ｍｇ／Ｌ＋ＫＴ１ｍｇ／Ｌ培养基上可诱导出愈伤组织并直接分化成苗,植株再生频率为３４％,最高达５０％。获得大豆高频率再生植株,收到较饱满的种子。     

从种子愈伤组织培养选育耐盐水稻突变体 1.愈伤组织诱导、植株再生及表观的植株性状变异

采用种子愈伤组织培养研究15个籼稻和硬稻品种的愈伤组织诱导及植株再生。还测定了选择性培养基不同氯化钠浓度对愈伤组织生长及植株再生的影响。不同品种愈伤组织诱导及植株再生的能力差异显著。在不加酵母及酪旦白提取物的培养基中,愈伤组织诱导频率从32％至85％。但当加入上述物质时,使诱导频率提高至将近100％。植株再生频率从IR8、IR26、IR30、IR36以及“Koshihikari”的0％至“台中65”的62％。植株再生频率随着愈伤组织培养时间的延长而下降。不过,“台中65”及“Nona Bokra”培养27周的愈伤组织还保持30％-40％的再生能力。“台中65“的愈伤组织即使生长了47周还能再生成植株。选择性培养基加进氯化钠对愈伤组织生长及随后的植株再生有显著影响。在氯化钠浓度为2％时,只有少数愈伤组织略有生长:氯化钠浓度低于1％时,愈伤组织生长良好。1.5％的氯化钠浓度抑制愈伤组织生长,但在第一筛选阶段仍然保持中等的生长速度。所有供试的品种,用培养11周的愈伤组织在氯化钠浓度较高的情况下植株再生频率仍较高。     

禾谷镰刀菌(Fusarium graminearum)毒素对小麦幼胚愈伤组织诱导和分化的影响

为了给普通小麦抗赤霉细胞工程育种中抗毒素细胞系选择技术体系的建立和方法的选择提供理论依据，研究了离体培养条件下禾谷镰刀菌毒素胁迫对小麦幼胚愈伤组织诱导和分化的影响。结果表明．禾谷镰刀茵毒素对小麦幼胚胚性愈伤组织的诱导具有低浓度促进和高浓度抑制、毒害作用，这种毒害作用降低了胚性愈伤组织诱导率和胚性愈伤组织质量，显著影响胚性愈伤组织再生频率。同时禾谷镰刀茵毒素胁迫降低了幼胚愈伤组织产量，并随胁迫浓度的升高，愈伤组织产量迅速下降。禾谷镰刀菌毒素对小麦幼胚培养特性的影响，表现出基因型间的差异，对感病基因型的毒害作用远大于抗病品种。     

橡胶树品种热研88-13易碎胚性愈伤组织的诱导及其植株再生

以高产优质橡胶品种热研88-13的幼嫩种子内珠被为外植体材料,对愈伤组织诱导培养基和继代培养基中的影响因素如植物生长调节剂的种类和浓度、钙离子浓度等进行了研究。经过连续6个月的继代选择培养,逐渐诱导出易碎的胚性愈伤组织,适宜的易碎胚性愈伤组织诱导培养基为:MH基本成分,添加0.5 mg/L KT,2.0 mg/L 2,4-D,11.0 mmol/L CaCl2,80.0 g/L蔗糖和2.0 g/L Phytagel。组织学切片证明长期继代培养的易碎胚性愈伤组织维持了胚性的状态。取继代培养2 a多的易碎胚性愈伤组织诱导体细胞胚的形成,得到了16 000多个体细胞胚。将这些胚进行进一步的转移培养,得到了115株再生植株。     

普通大麦(Hordum vulgare)未成熟胚愈伤组织的诱导和植株再生的初步研究

本文对五种基因型的普通大麦未成熟胚在离体培养下愈伤组织的诱导能力和植株再生能力进行了比较。初步结果表明:0.5—1.5毫米大小的未成熟胚为合适的培养材料,只要诱导培养基合适,各基因型都能诱导出愈伤组织,并达到较高的诱导率。但愈伤组织的质量在各基因型中存在显著差异。同时发现,再生植株诱导频率主要决定于愈伤组织的质量,而愈伤组织的质量直接由基因型和诱导培养基决定。不同的分化培养基在分化再生植株的作用上没有出现显著的差异。     

铁兰体细胞胚的诱导及植株再生

以铁兰无菌苗为外植体,研究不同激素配比对铁兰胚性愈伤组织诱导及体细胞胚形成的影响,并对体细胞胚的发生过程进行形态学观察.结果表明:2,4-D对铁兰胚性愈伤组织的诱导具有重要作用,诱导铁兰胚性愈伤组织的最适2,4-D浓度为2.0 mg/L;胚性愈伤组织在含ABA的培养基上可分化形成体细胞胚.形态及解剖学观察表明,愈伤组织既可以产生于外植体的外层细胞,又可产生于外植体的深层细胞.体细胞胚的形态发生经历了与合子胚形态发生相似的阶段,即经过球形胚、子叶形胚等.胚经过萌发、芽的伸长和生根后形成再生植株.     

茶树叶片组织培养的初步研究

用MS培养基附加以不同激素的种类及浓度的组合配比,探讨茶树叶片组织培养中对愈伤组织、胚状体、芽和小植株分化的能力。结果表明,连续用7、8号两组培养基作继代培养,新形成的愈伤组织都具有分化芽的能力。当这种愈伤组织转移到4号培养基上培养,可获得胚状体或胚性细胞团的分化,进而形成正常芽和苗。若将上述新形成的愈伤组织转移到9号培养基上培养,可使芽体有根的分化,从而得到健壮苗株。说明茶树叶片的组织培养中诱导愈伤组织,分化胚状体和小植株的形成所需要的外源激素的种类组合及浓度配比上存在明显差异。     

不同菜用大豆品种的胚胎发生与再生

以8个菜用大豆品种的未成熟子叶为外植体,在MS(附加高浓度2,4-D)培养基诱导愈伤组织形成、体细胞胚胎发生与再生植株.结果表明:八个菜用大豆品种的未成熟子叶均可诱导产生愈伤组织与体细胞胚胎发生,出愈率为14.71%～100%.体细胞胚胎发生率变化在0～36.76%之间,绿子叶菜用大豆没有形成体细胞胚.体细胞胚可正常萌发产生小植株,移栽成活结实.菜用大豆的出愈率和体细胞胚胎发生率与基因型有关.     

玉米再生体系建立及其影响因素的研究

以玉米自交系辽7980、丹9818、340、5026为材料,对影响成熟胚愈伤组织诱导、继代培养及分化的因素进行了研究。结果表明,2,4-D对于玉米成熟胚愈伤组织的诱导是必要的,基因型对胚性愈伤诱导的影响显著。在培养基中添加L-脯氨酸对提高胚性愈伤组织有显著作用,但对愈伤组织生长率没有明显影响。适量的BA和AgNO₃对植株的再生有促进作用。     

Comparative study of direct and indirect evaluations of frost tolerance in barley

The aim of this paper was to compare different field, laboratory and physiological methods for the evaluation of frost tolerance in barley; as well as to show both the possibilities and limitations of these individual methods and approaches. The tolerances of 39 barley cultivars and breeding lines were evaluated by four direct methods (based on the exposure of plants to frost) and three indirect (based on diagnostic traits and markers). The direct methods included the evaluation of: (1) field survival after five winters 1999–2004; (2) winter survival in a provocation pot test under natural conditions; (3) lethal temperature (LT50) of plants taken from a field in winter; and (4) LT50 of plants grown and hardened in a growth chamber. The indirect methods were based upon: (1) endogenous levels of abscisic acid (ABA); (2) water content (WC); (3) osmotic potential (OP) of the plants cultivated and hardened in growth chambers. All four direct methods correlated well with one another, and resulted in similar sequences of frost tolerance of the tested barley cultivars and lines. While it was only possible to distinguish individual barley cultivars based on field survival after the 2002/2003 winter, by using the provocation method it was possible to distinguish between barley cultivars after all of the winters. Moreover, the average winter survival of barley from multi-year pot tests was in correlation with the minimal LT50, evaluated in plants hardened in both the field or in growth chambers. From indirect indicators, the levels of ABA increased slightly, while WC and OP clearly decreased in all cultivars during cold hardening. No correlation was observed between LT50 and the ABA content. A significant correlation between LT50 and WC, and between LT50 and OP was found, but only after 1 or more weeks of cold acclimation of the plants had been completed, since both the duration and the rate of decline of these parameters differed in individual cultivars during the cold hardening. The LT50-values of barley cultivars not only accurately characterized their field tolerance to frost, but also were indicative of changes associated with the induction of frost tolerance in plants under the given conditions, when comparing the direct and indirect methods.     

Potato (Solanum tuberosum L.) cultivar response to bentazon and crop oil

Bentazon [3-(l-methylethyl)-(1H)-2,l,3-benzothiadiazin-4(3H)-one 2,2-dioxide] was applied at two rates to nine potato cultivars in the field, with or without a crop oil, to plant foliage at two different heights. Seven days after application of bentazon, foliar injury ranged from 0 to 36 percent depending on rate, cultivar, and plant height at the time of application. Of nine cultivars tested, Russet Burbank was the most susceptible to injury and Red LaSoda the least susceptible. A bentazon application rate of 1.12 kg a.i./ha, compared to a rate of 0.84 Kg a.i./ha, resulted in a biologically insignificant increase in foliar injury of two to four percent and a significant increase in weed control. The addition of crop oil to bentazon did not result in an increase of foliar injury, but increased the percentage of weeds controlled approximately two-fold. With one exception, application of bentazon to plants 8–15 cm in height resulted in less foliar injury than application to plants 20–30 cm in height. Application of bentazon to larger plants resulted in lower yield and quality. For most cultivars, the application of bentazon (1.12 kg/ha a.i.) and crop oil (2.3 L/ha) to 8 to 15 cm-tall potato plants resulted in no appreciable loss in potato tuber yield, while controlling 74–85 percent of broadleaf weeds.     

Performance of a High-Yielding Modern Rice Cultivar Takanari and Several Old and New Cultivars Grown with and without Chemical Fertilizer in a Submerged Paddy Field

Abstract

A high nitrogen-uptake capacity and effective use of absorbed nitrogen for dry matter and grain production are required to improve the production cost and environmental pollution. We characterized grain yield, dry matter production and nitrogen accumulation in six rice cultivars: Sekitori (released in 1848) and Aikoku (1882), referred to as SA cultivars hereafter; Koshihikari (1956); Nipponbare (1963) and Asanohikari (1987), referred to as NA cultivars hereafter; and Takanari (in 1990) as a high-yielding modern cultivar. The plants were grown with and without chemical fertilizer in a submerged paddy field. When plants were supplied with manure and chemical fertilizer, Takanari consistently produced the heaviest grain and dry matter, followed by the NA cultivars, and the SA cultivars the lightest. Dry matter production before heading was greater in Takanari and the NA cultivars due to the longer duration of vegetative growth. Dry matter production after heading was greatest in Takanari, with a larger crop growth rate (CGR), and smallest in the SA cultivars with a shorter ripening time. Greater dry matter production during ripening was accompanied by the greater accumulation of nitrogen by Takanari and NA cultivars. These plants developed a larger amount of roots. The smaller light extinction coefficient of the canopy was also attributed to the higher CGR in Takanari. When plants were grown without chemical fertilizer, Takanari also produced heavier grain and dry matter, followed by the NA cultivars. The heavier grain in these cultivars resulted from the greater dry matter production before heading, which was due to the longer period of vegetative growth. The greater dry matter production and nitrogen accumulation by Takanari and NA cultivars were evident when plants were grown with chemical fertilizer. Koshihikari was characterized by a higher CGR and greater nitrogen accumulation during ripening in the absence of chemical fertilizer which should be noted in efforts to decrease rates of nitrogen application.     

茶树氮吸收效率的早期鉴定技术研究

氮是植物生长的重要营养元素,在茶树栽培过程中常需施用大量氮肥,不仅消耗大量的资源,施用不当还会造成一系列环境问题。培育氮肥高效利用的茶树品种是解决这一问题的重要途径,而建立快速筛选高效株系的早期鉴定方法对于缩短育种茶树育种年限具有重要意义。本研究分析龙井43（LJ43）和中茶108（ZC108）两个茶树品种在不同氮素水平下对氨态氮和硝态氮的吸收与利用数据,通过与¹⁵N同位素标记技术的比对,验证非损伤微测技术（NMT）和实时荧光定量（qRT-PCR）技术在早期鉴定茶树株系氮素吸收利用能力方面的可行性与实用性,以期建立茶树氮吸收效率的室内早期鉴定技术。试验结果表明,¹⁵N同位素标记技术的稳定性和可重复性分别为89.51%、99.26%,而NMT的稳定性、可重复性分别为95.22%、96.76%;两种方法测定结果均显示茶树具有明显的喜铵特性;硝酸根转运蛋白基因CsNRT3.2和CsNRT2.4在两个品种中均表现出诱导上调表达效应,相比中茶108,龙井43中CsNRT2.4和CsNRT3.2具有更高的表达量,表明LJ43对外界氮源的响应高于ZC108。综上所述,认为NMT技术可在短时间内处理并测得茶树的瞬时吸收速率,且试验材料损耗少,可以用于茶树氮瞬时吸收速率的早期鉴定;CsNRT2.4和CsNRT3.2的表达量一定程度上反映了茶树对硝态氮吸收的能力。本研究可为氮高效茶树品种的早期鉴定技术建立提供依据。     

茶树对铬、镉的吸收积累特性研究

通过水培实验,研究了茶树(3个品种)受铬(Cr3+)、镉(Cd2+)毒害的表观症状以及铬、镉在茶树体内的分布、积累特性。结果表明:铬(20~100mg/L)、镉(1~5mg/L)胁迫下,茶树出现的毒害症状和对新梢生长的抑制效应因胁迫浓度、时间和品种而异。茶树各器官的铬、镉含量总体随铬、镉胁迫浓度的增加而增加,并且与胁迫浓度呈极显著或显著正相关。铬在茶树体内由高到低的分布次序是:吸收根>主根>主茎>枝条>老叶>新梢(一芽二叶);镉则为:吸收根>主根>主茎>新梢(一芽二叶)>枝条>老叶。根部吸收的铬、镉元素大部分被吸收根所固定,向地上部运输的比例较低,但镉在茶树体内的活性和迁移能力强于铬,运输到地上部的镉极易迁移到新梢中。新梢不同叶位镉含量的高低顺序为:第一叶>第二叶>第三叶>第四叶,新梢中的镉有明显向上富集的趋势。不同品种的茶树对铬、镉的吸收、积累和迁移能力有一定差异。     

利用CRISPR/Cas9敲除OsNramp5基因创制低镉籼稻

【目的】为了尝试快速培育低镉籼稻，【方法】选取广泛应用的杂交水稻亲本华占和五丰B以及常规品种五山丝苗和中早35为材料，通过CRISPR/Cas9技术创制OsNramp5基因突变株系，在镉污染及正常土壤中种植并测定突变株系籽粒（糙米）镉含量，其他相关元素含量亦同时在镉污染土壤种植条件下测定，在非镉污染土壤种植条件下考查OsNramp5基因敲除对农艺性状的影响。【结果】成功获得了前述品种的OsNramp5基因敲除株系；非镉污染条件下种植的4个品种OsNramp5基因敲除株系籽粒镉含量低于0.02 mg/kg，平均较野生型降低85.5%；而在镉污染土壤种植时，不同品种OsNramp5基因敲除株系籽粒镉含量低于0.1 mg/kg，平均比野生型降低94.8%；锰含量也降低52.7%，铬含量增加59.5%，铅含量在华占中增加79.1%，而在其他品种中无变化；铜、铁、锌、钙、硒和砷含量（后4种元素只在华占及衍生品系中检测）受影响较小或不受影响；OsNramp5基因敲除株系株高、结实率和千粒重较野生型小幅降低，而有效分蘖略微增加，产量平均减少6.9%。【结论】通过OsNramp5基因敲除，可以显著降低镉积累，但在某些种植条件下，代价为小幅产量损失；通过本研究获得的低镉OsNramp5基因敲除品系在镉污染地区具有较好利用潜力。     

利用CRISPR/Cas9敲除OsNramp5基因创制低镉籼稻

【目的】为了尝试快速培育低镉籼稻,【方法】选取广泛应用的杂交水稻亲本华占和五丰B以及常规品种五山丝苗和中早35为材料,通过CRISPR/Cas9技术创制OsNramp5基因突变株系,在镉污染及正常土壤中种植并测定突变株系籽粒(糙米)镉含量,其他相关元素含量亦同时在镉污染土壤种植条件下测定,在非镉污染土壤种植条件下考查OsNramp5基因敲除对农艺性状的影响【。结果】成功获得了前述品种的Os Nramp5基因敲除株系;非镉污染条件下种植的4个品种OsNramp5基因敲除株系籽粒镉含量低于0.02 mg/kg,平均较野生型降低85.5%;而在镉污染土壤种植时,不同品种OsNramp5基因敲除株系籽粒镉含量低于0.1 mg/kg,平均比野生型降低94.8%;锰含量也降低52.7%,铬含量增加59.5%,铅含量在华占中增加79.1%,而在其他品种中无变化;铜、铁、锌、钙、硒和砷含量(后4种元素只在华占及衍生品系中检测)受影响较小或不受影响;OsNramp5基因敲除株系株高、结实率和千粒重较野生型小幅降低,而有效分蘖略微增加,产量平均减少6.9%。【结论】通过OsNramp5基因敲除,可以显著降低镉积累,但在某些种植条件下,代价为小幅产量损失;通过本研究获得的低镉OsNramp5基因敲除品系在镉污染地区具有较好利用潜力。     

Comparison of vegetative development in soybean cultivars for low-latitude environments

Soybean production has been expanded into northern Brazil by the introduction of long-juvenile genes. These genes cause flowering to be delayed but specific changes in the sensitivity of plant development to temperature and photoperiod have not been resolved in these commercial cultivars. This study was undertaken to resolve possible changes in long-juvenile plants associated with rates of plant emergence, leaf appearance, and flowering. Eight cultivars, including three long-juvenile cultivars, were sown each week throughout a year in a plastic greenhouse so that the plants would develop under differing temperature and photoperiod. While there were differences among cultivars in the cumulative temperature required for plant emergence and the rate of leaf appearance, these differences were not necessarily associated with the long-juvenile trait. An extended duration to flowering was confirmed for the three long-juvenile cultivars but this delay was not associated with any difference in sensitivity to temperature and photoperiod. The trait that distinguished the long-juvenile cultivars was a much lower maximum development rate towards flowering than that found in the other cultivars. However, cultivar differences were identified among the long-juvenile cultivars indicating the possibility of further selection of genotypes to fit specific environments in low-latitude regions.     

Greenhouse techniques to identify field resistance to the brown planthopper,Nilaparvata lugens (Stål) (Homoptera: Delphacidae), in rice cultivars

Two seedbox screening techniques were compared to determine levels of resistance of rice cultivars to three brown planthopper, Nilaparvata lugens (Stål), biotypes. The commonly used standard seedbox screening test (SSST) where eight second-instar nymphs were infested on 7-day-old seedlings was compared with a modification (MSST) where four second-instar nymphs were infested on each of 20-day-old plants. In the SSST, the initial infestation killed the susceptible cultivars whereas in the MSST the F₁ progeny killed the susceptible cultivars. The cultivars ASD 11, Wagwag, Utri Rajapan, IR46 and Kencana were susceptible in the SSST but moderately resistant or resistant in the MSST, indicating an increase in the level of resistance with plant age. Feeding rate and population growth of N. lugens were the factors involved in the resistance of 30-day-old plants. The MSST provides a rapid method for identifying field-resistant cultivars in the greenhouse and can be used to identify field-resistant lines in a rice-breeding programme for stable resistance to N. lugens.     

遮光对光照敏感型新梢白化茶春梢化学成分含量的影响

以光照敏感型新梢白化茶树‘黄-2’、‘黄-8’和‘黄-13’为供试材料,在春季一芽一叶期采用30%透光度的黑色遮阳网覆盖遮光处理7天,HLPC法检测一芽二叶新梢咖啡因、儿茶素类、氨基酸类和色素类化合物含量。结果显示,遮光处理对茶叶咖啡因含量无显著影响;遮光对儿茶素类总含量的影响因品种而异,在品系‘黄-2’显著提高,而品系‘黄-8’和‘黄-13’则显著降低;遮光显著提高品系‘黄-13’的氨基酸总含量,降低品系‘黄-8’氨基酸总含量,但对品系‘黄-2’无显著影响;遮光显著提高β-胡萝卜素、叶绿素a、叶绿素b和新黄质的含量,显著降低紫黄质含量。具有热耗散作用的黄体素在光照敏感新梢白化茶品种的含量很低,是其容易受到强光胁迫伤害的原因。在强光照条件下,光照敏感型新梢白化茶叶片呈现黄色,主要是叶绿素含量显著降低,而不是因为胡萝卜素和叶黄素含量提高所致。根据这些研究结果,作者认为:在高温强光季节采取适度遮光,将有助于光照敏感型新梢白化茶提高叶片叶绿素含量,增强光合作用,进而提高茶苗生长势和抗性,有助于提高茶苗移栽成活率。