Sporophytic and gametophytic recurrent selection for improvement of partial resistance to Alternaria leaf blight in sunflower (Helianthus annuus L.)

Alternaria leaf blight, caused by Alternaria helianthi Hanf., is one of the most important diseases of sunflower causing significant yield losses in several tropical countries. Yet, so far, only partial resistance for the disease has been discovered in the germplasm through conventional sporophytic selection. Therefore, the main objective of the present investigation was to compare sporophytic and gametophytic recurrent selection with the aim to enhance the level of resistance to Alternaria leaf blight. The base population was synthesized by random mating three populations- two interspecific derivatives involving different species of Helianthus and one germplasm accession based on their partial resistance to disease incidence. The base population was subjected to 1-2 cycles of both sporophytic and gametophytic selection. The gametophytic selection was practiced by applying pathogen culture filtrate to the stigma and style one hour before pollination. The selection response was measured by scoring the percent disease index at flowering, 15 days after flowering, and at physiological maturity and by quantifying economic yield gain. A significant reduction in mean per cent disease index values and a gain in seed yield were observed for both the types of selection cycles, but more so for gametophytic selection. The populations improved through gametophytic selection appear to be more promising as the pollen selection allowed the selection of rare favorable allelic combinations that would hardly be detected at the sporophytic level. A combination of gametophytic selection and conventional sporophytic selection should be considered as an effective tool in population improvement programs to achieve higher levels of resistance in relatively short time.     

小麦花粉管生长途径及受精过程经历时间的研究

应用常规石蜡切片和荧光显微镜技术对小麦花粉管生长途径及受精过程相应时间进行研究。结果表明，授粉后，花粉随即萌发，2个精子进入花粉管，营养核留在花粉粒内，不久解体。花粉管进入羽毛状柱头分支结构的细胞间隙，继续生长于花柱基部至子房顶部的引导组织的细胞间隙内。随后穿过子房壁，在子房壁与外珠被之间的缝隙中向珠孔方向生长。自花粉萌发至花粉管长入珠孔大约需要1 h。自花粉萌发并到达柱头分支结构中，花粉管均具明显的绿色荧光；但在引导组织区以及子房壁与外珠被之间生长的花粉管几乎看不到荧光。授粉后1 h花粉管经珠孔及珠心表皮细胞间隙进入1个助细胞，并释放精子。授粉后2～6 h精卵融合，授粉后16 h合子第一次分裂。授粉后2～3 h精子与极核融合，授粉后4 h初生胚乳核第一次分裂。授粉后2～16 h，即精卵融合至合子分裂前为花粉管通道法转化合适的时间，采用花柱横切滴加法转入外源DNA。     

Effect of Abscisic Acid on Pollen Germination and Tube Growth of Maize Genotypes1

As a preliminary step to using gametophytic selection to obtain genotypes of maize, Zea mays L., differing for abscisic acid (ABA) content and/or sensitivity, the influence of ABA on the in vitro germination and tube growth of maize pollen and the presence of genetic variability for these traits was investigated. In a first experiment, pollen samples of three genotypes were germinated on substrates containing increasing amounts of ABA. Low ABA concentration (0.5 νM) significantly increased pollen germination and tube growth, while 500 νM ABA significantly decreased both traits, thus indicating that ABA can influence the growth of maize pollen in vitro. In a second experiment, eleven inbred lines were screened for the in vitro reactivity of their microgametophytes to ABA. The lines differed significantly for the response (% of control) of tube length, but not of pollen germination to ABA.     

A procedure suitable for in vitro pollen selection inBrassica oleracea

Summary Experiments to determine whetherBrassica oleracea pollen could effect fertilisation following incubation in liquid culture medium are reported. Pollen was incubated in vitro, collected by centrifugation and used to pollinate compatible pistils. While retaining viability and the capacity to germinate such pollen was unable to penetrate the stigma papillae. However, incubated pollen produced functional tubes following pollination of styles from which the stigmas had been removed. These tubes grew through the style to the ovary and viable seed was obtained. The potential application of this procedure in pollen selection is discussed.     

The progamic phase in Cichorium intybus L. Pollen tube growth in the style,incompatibility reaction and gametophytic competition

Summary Pollen germination and tube growth were studied following compatible, incompatible and pseudo-compatible pollinations in chicory. Pollen germination begins 3 minutes after compatible pollinations. The earliest pollen tubes reach the ovary 17 minutes later. Many of the later germinating pollen tubes are arrested and burst at the stigma papillae. In the transmitting tissue inhibitional effects due to negative interactions between pollen tubes are frequently observed. Complete self-incompatibility results in total inhibition of germination. In case of pseudo-self-compatibility, some pollen germinate but germination and stigma penetration are delayed and often result in pollen bursting. There is no self-incompatibility reaction in the transmitting tract but if the pollen tubes reaching this tissue are relatively numerous, negative interactions between them occur as after compatible pollinations. An hypothesis is presented which attributes the negative interactions between pollen tubes to the diffusion of a substance from the growing pollen tubes. This substance would also provoke pollen bursting on the stigma.     

美国梾木(Cornus spp.)花粉活力及萌发特性

为了了解美国梾木花粉活力及萌发特性,采用扫描电镜技术对13个美国梾木无性系花粉形态进行观测与方差分析,并采用荧光显微技术和TTC染色法对无性系2号花粉萌发特性及花粉活力进行观察。结果显示:(1)不同无性系间花粉大小差异不显著,但外壁纹饰差异较大,极面沟距的变异系数最高且遗传最为稳定。(2)花序开花率为75%时花粉活力最高,整花4℃条件下短期保存花粉活力较高。(3)授粉后,多数花粉可在柱头上正常萌发,72 h后花粉管即可进入子房。研究认为,花粉外壁及极面沟距可作为鉴定美国梾木无性系的形态指标,美国梾木花粉活力及萌发特性是抗逆及丰产杂交组合的重要保证。     

‘砀山酥梨’花柱可授性及蜜蜂授粉特性的研究

采用半离体培养法和联苯胺-过氧化氢法研究了‘砀山酥梨’花粉萌发特性和柱头可授性。通过对‘酥梨’的最适授粉时间以及7种授粉处理坐果率的研究,探讨其授粉特性。结果表明：开花后每天酥梨柱头活性最佳的时间是11时至15时,柱头最佳授粉时间为开花后第4~5天;‘酥梨’花粉最适萌发温度为25℃,花粉管生长最有利的温度为22~25℃,在相同温度下,花粉萌发率和花粉管长度随时间增加而增加。7种授粉处理‘酥梨’的花朵坐果率中,没有昆虫授粉的‘酥梨’坐果率为0.48%和0.44%,自然授粉和人工授粉坐果率分别为2.83%和13.05%,‘酥梨’ב雪花梨’组蜜蜂授粉的坐果率为32.95%,新鲜花粉和商品花粉坐果率分别为70.2%和13.05%。由此可见,‘酥梨’是异花虫媒果树,蜜蜂可以在‘酥梨’柱头活性最高时进行授粉,从而提高‘酥梨’的坐果率,同时通过对‘酥梨’花器官生理活性和坐果习性的研究,为‘酥梨’的授粉和生产提供基础资料。     

油松花粉离体萌发及萌发特征研究

为了加强油松有性杂交,为种子园育种研究奠定基础,对油松花粉离体萌发的适宜条件及萌发特征进行了系统研究。结果表明,油松花粉在无蔗糖培养基中仍具一定萌发能力,蔗糖的添加可显著促进花粉萌发,但高浓度蔗糖可抑制花粉管的生长;油松花粉管易形成分枝,蔗糖浓度与分枝花粉频率呈正相关;硼酸的添加在培养初期可有效加速花粉萌发,但过高浓度的硼酸对花粉萌发具抑制作用;Ca2+对花粉萌发的影响较小,不同浓度Ca2+处理均可获得较高的萌发率;培养温度对花粉萌发的促进作用明显,但温度的持续升高将抑制花粉萌发。综合萌发率、花粉管生长及分枝花粉管频率,油松花粉萌发的最适液体培养基组成为：2.00%蔗糖+0.075%硼酸+0.005% Ca2+,26℃下光照培养,72 h时萌发率可达(96.10±0.47)%。     

桃花粉活性检测和离体萌发特性的研究

果树花粉活力直接影响授粉、受精乃至座果。为了解果树品种的花粉生活力,给引种栽培和杂交育种工作创造有利条件,以桃花粉为试验材料,采用染色方法检测花粉的活性。试验结果表明,含有蓝墨水的花粉培养基,花粉萌发率可以达到64.67%,与对照相比差异不明显,因此该方法比较适合于快速检测花粉活性;固体培养基培养2个品种桃花粉,萌发率比液体培养基培养分别高25%和20%。另外温度、花粉密度、培养基中蔗糖及硼酸浓度都对花粉萌发率和花粉管长度有影响,在温度为24℃、1粒花药/100 μL、10%的蔗糖或100 mg/L的硼酸的条件下,均有利于花粉萌发和花粉管生长。     

甘薯组(Batatas Section)种间、种内交配不亲和特性研究

以甘薯及其近缘野生种为材料,研究甘薯组种内和种间不亲和特性,主要结果如下:1.无论种内或种间,交配不亲和性是存在的;2.种间不亲和表现特点依组合不同,表现为3种类型:(1)花粉不萌发或产生极短花粉管,柱头的乳突细胞有胼胝质反应。(2)花粉萌发延迟,花粉管生长缓慢、停滞。(3)花粉正常萌发,花粉管生长受阻。几乎没有观察到正常     

万寿菊花粉活力及柱头可授性研究

为探明造成万寿菊雄性不育系结实率低的原因,对万寿菊花粉活力及柱头可授性进行了研究。采用花粉离体萌发法研究万寿菊自交系V-01花粉萌发适宜温度、花粉活力日变化和适宜贮存条件;用联苯胺-过氧化氢法检测万寿菊雄性不育系S-261、S-17-06-29、S-0191不育株柱头的可授性;水溶性苯胺蓝染色法检测花粉在柱头的萌发情况。结果表明：（1）万寿菊V-01适宜萌发温度介于25~30℃之间;V-01花粉活力日变化趋势为先升高后降低,11-13时采集的花粉萌发率最高;4℃干燥贮存是最适宜的花粉贮存条件。（2）万寿菊柱头形态呈‘γ’状时有可授性,可授性可持续3天。（3）万寿菊花粉授粉到柱头上1 h内即可萌发,授粉后2 h,花粉细胞达到花粉管内。     

麻疯树花粉管荧光染色及生长过程的研究

花粉管的生长是完成植物受精的重要生理过程,观察花粉管生长首先需要对雌蕊组织进行软化透明。本研究探索适合麻疯树雌蕊结构的透明方法,并利用苯胺蓝染色技术,在荧光显微镜下观察授粉后麻疯树花粉管的萌发和生长过程。结果表明:麻疯树雌蕊组织可采用NaOH溶液软化透明,较优的条件是2 mol·L-1的NaOH,50℃恒温箱内软化3~4 h;麻疯树授粉2 h后花粉管开始萌发,10 h后花粉管穿过花柱基部,11 h后在子房中观察到花粉管。麻疯树授粉后,花粉管进入子房至少需要11 h,完成受精则需要更长的时间。     

The ovular incompatibility system in Gasteria verrucosa (Mill.) H. Duval

Summary The nature and process of ovular incompatibility in Gasteria verrucosa was studied. Using different crossing programs and following the pollen germination and ovule penetration histochemically, an ovular gametophitic incompatibility system based on two or more loci is affirmed. Pollen tubes after self-and cross-pollination penetrate the micropyle. Although the percentage of seed setting reached about 60% after cross pollination no clear differences could be seen between the germination and ovule penetration of cross and self pollen.In the ovules, multiple penetration of micropyles and a preference for the first position ovules was observed. Results of semi-compatible crossings and mixed crossings indicated a gametophytic two or more loci incompatibility system. Ovular incompatibility was manifested by the degeneration of the sporophytic ovular tissues. In Gasteria verrucosa the system of ovular incompatibility does not seem to be restricted to the ovule.     

珍稀濒危物种大别山五针松花粉离体萌发的研究

为了研究大别山五针松濒危的原因,并为人工杂交育种和保护该物种提供理论依据。本试验采用花粉离体萌发法,设置正交试验,研究了蔗糖、硼、钙对其萌发率及花粉管生长的影响。结果表明：蔗糖是影响大别山五针松花粉萌发和花粉管生长的关键因子,其次是硼酸,氯化钙对其影响最小;蔗糖的添加,明显促进花粉萌发,但高浓度蔗糖花粉管的生长有很大的抑制作用;适当添加硼酸可以促进花粉萌发,但不利于花粉管的生长;氯化钙对花粉萌发影响最小,不同浓度的氯化钙均可以获得较高的萌发率。综合萌发率、花粉管生长,大别山五针松花粉萌发的最适培养基为：4%蔗糖+0.05%硼酸+0.025%氯化钙。     

Increasing resistance to Fusarium crown and root rot in asparagus by gametophyte selection

In garden asparagus, Fusarium crown and root rot is the main cause of crop decline. Since chemical treatments are inefficient, efforts should focus on the development of resistant cultivars to control the disease. Toxic culture filtrate (TCF) of F. oxysporum has affected asparagus pollen germination and tube growth. Consequently, gametophyte selection was evaluated to ascertain if the application of selective agents at this level could increase selection efficiency. Two susceptible pistillate plants and one tolerant and one susceptible staminate plants were used in controlled crosses. Before pollination, a drop of a germination vehicle with TCF or without it was applied to the stigmas. Some pollinated pistils were fixed and analyzed by fluorescence microscopy; the rest were left on the plant for seed production. Fifty to 200 seeds were obtained per treatment combination (staminate plant x pistillate plant x pollination vehicle). The derived plantlets were inoculated in vitroand evaluated for disease symptoms. The application of TCF to stigmas reduced pollen germination and tube growth compared with untreated controls,regardless of the genotypic combination. Pollen germination and tube growth was poorer for the tolerant staminate genotype than for the susceptible one. When the TCF was applied, the number of seeds per pollination in comparison with the controls diminished only when the susceptible genotype was the pollinator. The percentage of affected root area of the progenies obtained after applying the TCF was lower than in the controls only when the tolerant genotype was the pollinator. Increasing Fusarium resistance in asparagus by means of gametophyte selection seems feasible. This revised version was published online in August 2006 with corrections to the Cover Date.     

华北驼绒藜花粉萌发特性及不同授粉方式对其结实率的影响

为寻找华北驼绒藜结实率低、群落退化的原因,在栽培条件下对其花粉生活力、寿命、柱头萌发率与结实率的关系进行了相关性研究。采用同株、异株、自然授粉处理华北驼绒藜,同株授粉结实率(81.53%)显著低于异株授粉(88.6%)和自然授粉(92.6%)的结实率;采用TTC法对华北驼绒藜花粉寿命和生活力的研究结果表明,其平均寿命为6d,最佳授粉时间为1~3d,生活力达95%左右,不存在花粉败育和短命的问题,即不存在小孢子发育障碍。3种授粉方式处理后,采用荧光显微镜观察华北驼绒藜花粉在柱头上的萌发率,萌发高峰集中在授粉后3~4h,萌发率达90%以上,三者不存在显著差异,即华北驼绒藜不存在花粉与柱头的识别障碍。结实率与花粉在柱头萌发率的相关分析结果表明,二者不存在相关性。研究初步认为,同株授粉产生结实率低的原因可能发生在授精或胚胎发育过程,有待进一步研究。目前,华北驼绒藜野生群落严重退化,植被密度减少,可能存在大量自株授粉是造成群落退化的原因之一。     

植物生长调节剂对‘秦冠’苹果花粉管生长和花粉萌发的影响

研究吲哚乙酸、萘乙酸、赤霉素、2,4-D、6-BA5种植物生长调节剂对苹果花粉管生长和花粉萌发的影响。以贮藏6个月的‘秦冠’花粉为材料,配制不同浓度梯度植物生长调节剂的固体培养基培养花粉,研究不同浓度植物激素对花粉管生长和花粉萌发的影响。结果表明,当吲哚乙酸≤1 mg/L、萘乙酸≤1 mg/L、6-BA≤5 mg/L、赤霉素≤10 mg/L时对花粉萌发有轻微促进作用,当浓度增大时抑制作用较突出;2,4-D对花粉萌发存在明显抑制作用;当赤霉素、2,4-D、吲哚乙酸、萘乙酸、6-BA浓度分别达到50、30、30、15、25 mg/L时,花粉遭受浓度迫害出现畸形扭曲。吲哚乙酸、萘乙酸、赤霉素、2,4-D、6-BA对‘秦冠’花粉管生长和花粉萌发均无明显促进作用,并且随浓度增大抑制作用较明显,人工辅助授粉时用于添加剂效果不理想。     

新疆‘晚丰’巴旦木花粉萌发培养基组分的筛选

为提高人工授粉中巴旦木的产量,本研究采用花粉离体培养法,筛选新疆‘晚丰’巴旦木品种花粉萌发中不同培养基组分的配方施用量,研究不同培养基配方组分对花粉萌发的影响。实验中在培养基内添加尿素、硼酸、氯化钙、吲哚乙酸、赤霉素和蔗糖,均能促进花粉萌发和花粉管生长,在一定范围内,花粉萌发率和花粉管长度均随培养基配方组份量的增加而增加,但超过一定添加量时,促进作用逐渐减弱并出现抑制作用。研究结果显示:添加0.1%尿素、0.01%硼酸、0.005%氯化钙、0.0001%吲哚乙酸、0.0015%赤霉素和10%蔗糖时,新疆‘晚丰’巴旦木品种花粉萌发率最高,花粉管长度也达到最大值。本研究结果为实践生产中人工授粉和相关研究提供参考和理论依据,也为进一步顺利进行亲和授粉研究提供前期理论基础。     

Storage of avocado pollen

Summary Avocado pollen was stored at a range of temperatures and relative humidities (RH) for up to one year and the pollen was tested for viability in vivo.Pollen stored for one month was capable of germination on the stigma and penetrating the ovule when stored at 4°C with <1,23,55 and 75% r.h. and at -196°C with 0% r.h. Most pollen samples stored at 25 and -15°C at a range of r.h. would germinate on the stigma but none would penetrate the ovule.After one year of storage, pollen at 4°C and <1 and 23% r.h. would germinate on the stigma but would not penetrate the ovule. There was no germination of pollen stored at 4°C and 55 and 75% r.h. Only pollen stored at -196°C and 0% r.h. would penetrate the ovule, but thawing and refreezing once during the year destroyed the viability.     

Pollen viability and stigma receptivity in <Emphasis Type="Italic">Lilium</Emphasis> during anthesis

Pollen viability and stigma receptivity are prerequisites for successful pollination and seed set in flowering plants. In this study, the pollen viabilities and stigma receptivities of nine Lilium genotypes (six cultivars and three species native to China) were assayed by in vitro pollen germination and the benzidine-H₂O₂ method, respectively. Embryo sac development during anthesis was observed to further ensure the timing of controlled pollination. In addition, the relationship between stigma secretion and stigma receptivity was studied to estimate the pollination time based on phenotype. Anthers cracked on the day of flowering in all genotypes, but pollen germination during anthesis was not observed in Asiatic hybrids excepted for ‘Tiny pudhye’, which exhibited low pollen viability for a short period of time (from 0 to 1 day after anthesis). In the other genotypes, pollen germination rates were highest on anthesis (five of seven genotypes), 0–1 day after anthesis (L. sulphureum), or 0–2 days after anthesis (one Longiflorum hybrid), and then gradually decreased with days after anthesis. While, stigma receptivity first increased and then decreased during anthesis. For most genotypes, stigmas began to be receptive 1 day after anthesis, and all genotypes exhibited stigma receptivity at 2 days after anthesis. The durations of stigma receptivity and strongest stigma receptivity, were genotype dependent, and were 5–8 days and 1–4 days, respectively. Moreover, on the first flowering day, 6 of 7 genotypes had mature embryo sacs, and at the time at which stigmas began to be receptive, all tested genotypes had mature embryo sacs. Some Lilium genotypes showed stigma secretion, which can be a sign of stigma receptivity. Stigmas became receptive and reached highest receptivity within 1 day of the first appearance of secretion on the surface of the stigma and at peaking, respectively. The results of this study are valuable for the implementation of successful Lilium breeding programs.