玉米小孢子高频再生培养技术程序研究

以大田种植的玉米供体植株为实验材料,以提高游离小孢子存活率及成胚率为重点进行了技术优化研究。摸索出了一些行之有效的技术措施,提高了游离小孢子的离体培养反应,获得了一批再生植株。通过技术优化,建立玉米小孢子离体培养高频再生技术程序,在此基础上构建了玉米DH育种技术平台。应用这一技术平台,可以在当年获得玉米DH纯系的种子,第二年即可安排大田纯系鉴定。     

轮作对甘薯地线虫群落组成及多样性的影响

针对甘薯茎线虫病严重的多年连种区进行轮作玉米试验,研究连种甘薯、轮作玉米对甘薯地土壤线虫群落种类组成、垂直分布、线虫群落的多样性的影响,其结果共获得3目8科15属线虫。连种甘薯地极优势属为茎属Ditylenchus,甘薯地轮作玉米其极优势属为头叶线虫属Cephalobus、真头叶线虫属Eucephalobus,滑刃Aphelenchoides和真滑刃线虫属Aphelenchus。连种甘薯和轮作玉米地同一土层线虫属数量间没有显著差异,线虫数量间有显著差异;应用Shannon多样性指数、J均匀度指数、SR丰富度指数分析了连种甘薯和轮作玉米地土壤线虫群落的多样性,其中Shannon指数、SR指数最敏感,应用WI指数分析了土壤线虫群落的营养结构,WI指数可反映轮作玉米与连种甘薯间土壤线虫群落结构的差异;轮作可减少植物寄生线虫种类和数量,增加食细菌和食真菌线虫的数量,可改变线虫群落的营养结构。     

甘蓝型油菜小孢子培养技术及其单倍体应用研究进展

油菜小孢子培养已经成为油菜育种和生物技术的一项重要内容.由于影响小孢子成胚和成苗的因素较多,低诱导频率及成苗率严重限制了该技术在育种中的应用.为了全面了解小孢子胚状体发生能力及影响再生体系的各项因素,掌握该技术的研究动态和发展方向,总结了近20年来国内外影响小孢子培养的主要因素:供体植株基因型、供体植株生长条件及生理状态、小孢子发育时期、培养基组成、培养条件,及其在双单倍体育种、诱变育种、转基因育种等领域中的应用.     

活性炭对小白菜游离小孢子培养的影响

以24个小白菜品种为材料,通过在NLN培养基中添加不同浓度活性炭,研究其对游离小孢子培养的影响.结果表明:在NLN-13培养基中添加活性炭(0.5 g/L),有利于小孢子胚的诱导和形成,供试材料(N4,N8)添加活性炭处理胚诱导率较未添加活性炭的处理分别提高19,66.5倍.采用添加活性炭(0.5 g/L)的NLN培养基对24份小白菜品种进行游离小孢子培养,有20份材料诱导出胚,培养成功率为83.3%.表明添加活性炭的培养基对小白菜游离小孢子培养有较好的效果.不同基因型间胚诱导率差别较大,每花蕾诱导胚数为0.4～86.7个.依据胚诱导率的高低可将其分为极易诱导、易诱导、难诱导和不能诱导出胚4类.B5 6-BA 0.2 mg/L NAA 0.02 mg/L 活性炭0.5 g/L 3%蔗糖 1%琼脂培养基有利于幼胚长成植株.     

提高大白菜游离小孢子胚诱导率的研究

采用游离小孢子培养方法,对提高大白菜小孢子胚诱导和形成的因素进行了研究。结果表明,24个大白菜品种（品系）在NLN-13液体培养基中有21个品种诱导出胚。各品种间小孢子胚产量差别极大,每花蕾产胚在0．1-14．66个之间。比较添加不同浓度蔗糖和激素的NLN培养基的培养效果,以NLN＋13％蔗糖的效果较好。接种后,小孢子在33℃高温预处理24—48h,产胚量较高。B5＋6．BA0．2mg／L＋3％蔗糖＋1％琼脂培养基有利于小孢子胚长成植株。     

提高油菜游离小孢子胚诱导频率的研究

以11个甘蓝型油菜基因型为材料,采用NLN培养基进行游离小孢子培养,对如何提高可培养的基因型范围和产胚率进行了研究。结果表明,11个油菜基因型中有10个基因型可以诱导出胚,培养成功率达90.9%,表明采用NLN培养基进行游离小孢子培养油菜基因型范围比较宽,但各基因型间小孢子胚产量差别很大,每花蕾产胚量为0.08~3.53个,TR4和TR9两个基因型每花蕾产胚可达3.23,3.53个。以TR4和TR9两个基因型为试材,进一步改进培养基和培养方法,采用NLN培养基中添加激素和活性炭方法,可大大提高产胚率,产胚量分别达到7.11和10.05个/蕾;接种后,小孢子经33℃高温预处理可显著影响产胚量。子叶形小孢子胚在光下适当培养后转入B5 BA 0.2 mg/L NAA 0.02 mg/L继代培养基上,大多数胚能长成绿芽,B5 6-BA 0.2 mg/L 3%蔗糖 1%琼脂培养基有利于小孢子胚长成植株。     

大田油菜游离小孢子培养高频胚状体诱导及植株再生

以大田生长的双低油菜品系“9841”为供试材料，研究了影响小孢子培养反应的因素。结果表明：始花期前、后一周的主轴花序为起始材料经1-2d4℃低温预处理，B5培养基为提取液游离小孢子培养于含脯氨酸200mg/L、谷氨酰胺800mg/L、蔗糖13%的NLN培养基，32℃热激1d后转入25℃常温静置培养，二周后进行45rpm振荡培养，可提高胚状体的诱导频率及正常植株再生率。     

小麦游离小孢子培养的研究

在游离小孢子培养过程中认为子房共培养可显著地提高游离小孢子胚状体愈伤率和再生率 ,子房共培养以 2 0个子房为宜。低温子房共培养胚状体发生和愈伤率低 ,但绿苗分化率较高 ,施加PAA、谷氨酰胺对游离小孢子形成的胚状体有促进作用。麦芽糖是游离小孢子培养最好的碳源 ,以蔗糖、葡萄糖以及葡萄糖和果糖搭配作碳源均未获得胚状体。检查培养 3d后的小孢子活力发现 ,唯有麦芽糖作碳源培养的小孢子有活力 ;认为高浓度的蔗糖、葡萄糖以及葡萄糖加果糖对小孢子有毒害作用。当葡萄糖浓度达到 2 0mmol/L时就对小孢子有毒害作用且致死。当在麦芽糖培养基中培养 1d后加入高浓度的葡萄糖发现小孢子存有活力 ,而且随着起始培养天数的增加活力也增加。对Y77品系来讲 ,适宜小孢子培养的麦芽糖浓度为 0 12mol/L。饥饿和热激 (33℃ )预处理对游离小孢子培养有促进胚状体和愈伤形成的作用     

利用小孢子培养技术创制红菜薹早熟DH系

为获得早熟型的双单倍体(DH)植株,加快早熟型红菜薹育种进程,本研究以15个早熟型红菜薹品种为试材,进行小孢子培养,观察小孢子发育过程并对再生植株进行倍性鉴定。结果表明,当花瓣/花药(P/A)长度比值为0.6～0.8时,红菜薹小孢子72%～78%处于单核靠边期;共有6个基因型红菜薹出胚,其中出胚率最大的为gy21-55,平均出胚率为14.3胚/蕾;选取子叶形胚进行植株再生,成苗率在90%以上;利用流式细胞仪对再生植株进行倍性鉴定,平均自然加倍率达76.8%。gy21-40、gy21-55表现出早熟特性,与对照植株相比获得的DH植株可分别提早5 d和8 d抽薹。     

影响油菜游离小孢子培养再生因素的研究进展

7胚的培养及植株再生 7．1培养方式 经过高温处理后的小孢子，在24-25℃黑暗条件下静置培养2-3周，并保持良好的通气状况，促进胚的形成。胚龄是指从接种到胚被转移至固体培养基上，这段时间的长短。诱导植株再生最适合的胚龄是27d，0．2-0．3cm大小的胚比0．6-0．8cm的胚好。当肉眼可见胚状体（球状至鱼雷状早期）后，可以采用以下两种方法进行培养：     

影响玉米花药培养效率的因素研究

对影响玉米花粉胚诱导的因素进行了研究。结果表明 :花药接种前对雄穗进行 5℃低温预处理 15～ 2 1d效果较好 ;接种后高温 (33℃ )连续处理花药 ,胚出现早、质量好且效率高 ;活性炭 (0 5 g/L)加入培养基中能显著地提高胚诱导效率。此外 ,还对基因型间差异进行了比较分析     

Radiation Balance Components of Maize Hybrids Grown at Various Plant Densities

Components of the energy and heat balances were examined in two maize hybrids grown at three different plant densities (40, 70 and 100 thousand plants per hectare). One of the hybrids was drought tolerant, while the other was bred for cultivation under irrigated conditions. An increase in plant density influenced not only the size of the leaf area, but also the distribution of the leaves at various plant heights. The extinction coefficient, which provides a quantification of radiation penetration, was higher in the irrigated treatments. By contrast to the other two treatments, the plant canopy in the thinly sown stands remained open throughout the vegetation period, and thus behaved quite differently to the closed stands, making it impossible to compare them. Smaller albedo values were recorded for the hybrid bred for irrigation and in thinly sown stands. The low plant density allowed more energy to reach the soil, from which it was reflected, making a considerable contribution to the final temperature in the stand. The latent heat, in keeping with the quantity of water transpired, was the greatest in the densely sown stands. There was little difference between the latent heat values of the normal and dense stands in either hybrid, indicating that they both had a similar sensitivity to increased stand density. If sufficient water is available it would appear that the stand density could be increased even for the drought-tolerant hybrid.     

玉米各亚种的核型研究

研究了玉米８个亚种，２个亚型的核型。所有材料的根尖细胞染色体数目均为２ｎ＝２０，主要由中部和近中部着丝点染色体组成，第６对染色体短臂均具随体。但臂比值不同，可区分为Ａ１（ｍ染色体）、Ａ２（ｓｍ）、Ａ３（ｓｔ）三类。在核型中具中部着丝点染色体的数目，罕见栽培或原始类型多于广泛栽培的类型。玉米各亚种的核型进化的趋势是由对称向不对称方向发展。     

2种甜玉米自交系幼胚再生体系的建立

本试验研究不同基因型、幼胚大小、培养基类型及培养基添加物对2种甜玉米(2-105和2-118)自交系幼胚愈伤及胚性愈伤组织诱导率的影响。结果表明,基因型、培养基类型、幼胚大小及不同浓度脯氨酸和2,4-D处理对愈伤及胚性愈伤诱导率影响差异均显著。2种基因型最适宜的基本培养基为N6培养基,且2-118的愈伤及胚性愈伤诱导率显著高于2-105。幼胚在1.5~2.0 mm时,2-105和2-118的愈伤及胚性愈伤诱导率均最高,分别在77%和51%以上。2-105和2-118在脯氨酸浓度分别为650 mg/L、750 mg/L时幼胚的愈伤及胚性愈伤诱导率最高,且显著高于其它2个水平。2种基因型在2.5 mg/L的2,4-D诱导下愈伤及胚性愈伤诱导率均最高,且均显著高于其它2个水平。本研究获得了2种甜玉米自交系的再生体系,为甜玉米遗传转化和分子育种提供基础。     

Effects of Simulated Hail Damage on the Yield of Forage Maize

Hail damage was simulated with defoliation, leaf tearing and damage to ears and stems, applied at five stages and three levels of intensity on forage maize grown in Belgium. Leaf tearing alone had little effect on total dry matter yield. Decrease of yield was the highest when treatments were applied around tasseling-anthesis. Effects on total dry matter content were small.     

玉米CMS-S小孢子败育过程中的细胞程序性死亡

以玉米（Zea mays L.）S型细胞质雄性不育系(CMS-S)的一对近等基因系S-Mo17^Rf3Rf3 和S-Mo17^rf3rf3为材料，采用TdT介导的dUTP DNA末端标记（TUNEL）、细胞色素C免疫原位杂交和DNA寡聚核小体片段电泳等方法，分别在细胞学水平和DNA水平上研究了玉米CMS-S小孢子败育的细胞程序性死亡（PCD）过程。结果表明，在花粉母细胞减数分裂后的四分体解离时期，不育花药的绒粘层细胞较可育花药提前裂解；在不育系S-Mo17^rf3rf3花药和花粉S-rf3中均明显出现PCD过程的DNA片段化以及线粒体细胞色素C外渗的现象，证明了玉米CMS-S的花粉败育与花药绒粘层细胞的提前凋亡和小孢子细胞的程序性死亡有关。     

Growth Patterns of Early Maturing Maize Cultivars

Growth patterns of two early maturing maize cultivars were analyzed, harvesting shoots twice weekly in 1983. Until the end of leaf expansion the leaf dry matter accumulation was highly synchronized in both hybrids. After a short cessation leaf dry weight accumulation was resumed at low to medium rates during the main part of August, during September losses prevailed. During these two months fluctuations in rates were frequent and synchronisation between genotypes small. For stems no transitory cessation of dry matter accumulation occurred; synchronisation between genotypes and fluctuation in rates were less than observed for leaf dry matter accumulation. Losses in stem dry weight mostly coincided with rapid ear growth. Ear dry weight accumulation steeply increased during the first three weeks of August. Fluctuations in rates were large during the following 30 days of rapid growth. This period ended by a sharp decline in rates. Phosphofructokinase activity of upper leaves was maximum in early July followed by a first minimum shortly before anthesis; thereafter the average activity did not change much. The hybrid with higher initial shoot growth rates had a higher phosphofructokinase activity at the same time.     

Effects of Nitrogen Fertilization on Yield and Yield Components of Tropical Maize Cultivars

Six tropical maize cultivars were grown in Thailand on a reddish brown latosol under three nitrogen regimes, 0, 40 and 80 kg N/ha. With regard to biomass and economic yield several "cultivar by N fertilization response types" were discriminated. The low fertilization type was represented by an early and a late maturing cultivar, both yielded comparatively well without additional N and responded little to N fertilization. An opaque 2 cultivar and a commercial hybrid belonged to the high fertilization type, they had a very low yield without N and needed high N fertilization to produce a high yield. The other genotypes were of an intermediate type with comparatively high yields without N, a considerable increase in yield at 40 kg N/ha and some increase from 40 to 80 kg N/ha. Genotypic variability for the harvest index was high but values were at the usual low level of tropical cultivars. High fertilization types reacted to an additional N supply with a marked increase in kernel number/plant and little increase in kernel weight. The early maturing low fertilization type reacted quite the opposite way. All other genotypes predominantly increased kernel number from 0 to 40 kg N/ha and kernel weight from 40 to 80 kg N/ha.     

新型植物生长调节剂B2对玉米光合荧光特性及产量的影响

为探讨新型植物生长调节剂B2拌种和叶面喷施对玉米农艺性状、光合特性、叶绿素荧光参数及产量的影响,明确B2拌种和叶面喷施的适宜剂量,以德美亚1号为材料,采用田间随机区组试验方法,设置B2拌种剂量0（CK）、25（BT1）、50（BT2）和100（BT3）mg/kg,在6展叶期叶面喷施剂量0（CK₁）、25（YT1）、50（YT2）和100（YT3）mg/L。结果表明：与CK和CK₁相比,B2拌种和叶面喷施处理均能降低玉米株高和穗位高,增加玉米茎粗、叶面积和干物质积累量;提高玉米叶片SPAD值、净光合速率（P_n）和蒸腾速率（T_r）;提高实际量子产量Y(Ⅱ)、相对电子传递效率（ETR）、最大光化学效率（F_v/F_m）和潜在光化学活性（F_v/F_o）,降低非光化学猝灭系数（NPQ）;B2拌种和叶面喷施处理产量较CK和CK₁分别增加5.90%、14.20%、5.81%和11.72%、14.40%、5.36%。综上所述,B2拌种剂量为50mg/kg、叶面喷施剂量为50mg/L时,可显著增加玉米产量。