水稻胚乳细胞发育的结构观察及其矿质元素分析

 为了探明水稻胚乳发育的过程和糊粉层形成的机理, 用光学显微镜和电子显微镜观察了水稻糊粉层细胞和内胚乳细胞在颖果发育过程中的结构变化,用能谱仪分析了胚乳细胞中元素的种类和相对含量。结果表明, 糊粉层细胞是由胚乳表层细胞转化而来的。糊粉层细胞中的P、K、Mg和Ca等矿质元素含量要明显高于内胚乳细胞。发育初期糊粉层细胞中富含线粒体、圆球体和小液泡;发育中后期小液泡积累蛋白质和矿质元素而形成糊粉粒。在发育中后期,内胚乳细胞随着细胞内淀粉体的充实,细胞核发生形变而衰亡;而糊粉层细胞的核在发育过程中不消亡。糊粉层的形成与表层细胞积聚矿质和脂类等“灌浆废物”(指非内胚乳细胞的贮藏物)有关。因而,转运灌浆物质多的胚乳背部,其糊粉层细胞的层数要比腹部和侧部多。谷物胚乳发育分为游离核期、细胞化期、分化期和成熟期四个时期。     

小麦乙烯响应转录因子基因 wfzp突变体颖果发育的研究

小麦WFZP基因编码的乙烯响应因子是植物中特有的转录因子，参与果实成熟、花叶器官衰老、脱落和种子发育等过程。为探明 wfzp基因突变对小麦颖果发育的影响，以甲基磺酸乙酯诱变的两个 wfzp基因突变体（ wfzp1和 wfzp2）和野生型小麦品种济麦22为材料，运用树脂超薄切片和显微观察技术观察小麦WFZP基因突变体颖果的形态结构发育特征。结果显示， wfzp突变体颖果发育有如下特征：（1）在颖果发育早期，两个突变体的外果皮发育进程较快，中果皮降解时间提前； wfzp1内果皮管细胞消失较快，与 wfzp1突变体相比， wfzp2突变体的管细胞发育缓慢；（2）两个突变体胚乳细胞的分化和发育明显较快，细胞中淀粉和蛋白体的积累量明显增多，细胞的充实度高，尤其在 wfzp2突变体中表现较为明显；（3）两个突变体胚乳传递细胞与糊粉层传递细胞的生长分化进程加快。研究结果为进一步探究乙烯响应转录因子调控小麦颖果发育提供了形态学参考。     

春季低温对小麦颖果发育的影响

为了探明春季低温（倒春寒）导致小麦减产的细胞学机理，以扬麦15为材料，在小麦幼穗处于雌雄蕊分化期至药隔期之间时，设置-3 ℃低温处理，通过树脂半薄切片对颖果不同发育时期的形态结构进行观察。结果表明：（1）在颖果发育早期，低温促进了颖果的长度生长，抑制了宽度生长；与对照相比，低温处理组小麦颖果发育进程缩短；（2）低温处理组颖果果皮发育早，中果皮降解速度快；（3）低温处理增加了胚乳淀粉体的积累量，降低了蛋白体的积累量，发育后期胚乳充实度显著低于对照组；（4）与对照相比，低温处理后颖果韧皮部面积和珠心突起传递细胞面积增大。以上结果表明，春季低温处理更有利于养分运输组织的发育，使得灌浆时间缩短，最终导致淀粉积累量增加及颖果早熟。     

扬麦16“灌浆快”与颖果显微结构和内源生长素的关系

扬麦16是长江中下游麦区种植面积最大的品种,具有灌浆速度快、粒重高等特点。为探明扬麦16"灌浆快"与颖果结构和内源生长素的关系,以宁麦13为对照,显微观察了扬麦16颖果维管束、胚乳细胞和传递细胞发育的变化特征,测定了千粒重和生长素含量。结果表明,扬麦16胚乳中淀粉体和蛋白体发育好,胚乳充实饱满,最终粒重高;灌浆过程中扬麦16颖果内源生长素水平高,促进了同化物向籽粒的调运和胚乳细胞的分裂;扬麦16颖果维管束发达,筛管面积大,韧皮部卸载能力强;扬麦16颖果传递细胞分化早,细胞壁内突结构发达,养分转运能力强。     

水稻胚乳组织的结构观察

【目的】阐明水稻糊粉层细胞、亚糊粉层细胞与中心胚乳贮藏细胞的结构特性。【方法】采用光镜、透射电镜与扫描电镜对水稻胚乳组织进行观察研究。【结果】糊粉层细胞分化过程中,大液泡变成小体积蛋白贮存液泡,蛋白贮存液泡又转变成糊粉粒。颖果背部比腹部有更多层糊粉层,但背部糊粉层细胞内糊粉粒的形成与积累速度却较慢。亚糊粉层细胞起初含有一些脂质体,后来脂质体消失,而其内部淀粉体与蛋白体逐渐增多。中心胚乳贮藏细胞含有淀粉体与蛋白体,蛋白体以液泡型蛋白体为主,它们可以相互融合而变大。中心胚乳贮藏细胞内的淀粉积累速度明显快于亚糊粉层细胞内的。成熟颖果的中心胚乳贮藏细胞内淀粉体最为密集,背部和侧部的亚糊粉层细胞内淀粉体排列较疏松,腹部的亚糊粉层细胞内淀粉体最为稀疏。【结论】水稻颖果背部与腹部的糊粉层细胞和亚糊粉层细胞的结构差异可能与养分吸收与转运有关;中心胚乳贮藏细胞内淀粉体发育速度快于亚糊粉层细胞。     

四个不同粒重水稻品种颖果发育的比较

 以粒重差异较大的4个水稻品种为供试材料,采用树脂切片、酶解胚乳细胞和显微观察等方法,比较研究了品种间在颖果生长、胚乳细胞增殖、果皮和胚乳结构等方面的差异,探讨了影响颖果生长的因素。 大粒品种颖果发育时间较小粒品种长,其胚乳细胞数、胚乳干质量及单个胚乳细胞平均干质量均高于小粒品种。在粒重相近的情况下,籼稻颖果发育和淀粉积累快于粳稻。与小粒品种相比,大粒品种子房壁细胞中淀粉粒多,子房壁细胞生长的持续时间长,果皮及背部维管束衰亡迟。 小粒品种胚乳外层细胞在花后7 d已转化成糊粉层细胞,大粒品种胚乳外层细胞要在花后10 d才转化成糊粉层细胞。 大粒品种的库容大和生理活性期长是其颖果能显著增大的生理原因。     

小麦内胚乳细胞及其淀粉体的发育

为了探明小麦内胚乳细胞发育和淀粉体生长的规律,以不同发育天数的小麦颖果为材料,利用树脂半薄切片和胚乳细胞分离等方法研究了内胚乳细胞及其淀粉体的形态、数目和生长过程.结果如下:(1)小麦内胚乳细胞呈不规则的多面体形态,内含大、小两种淀粉体,大淀粉体呈鹅卵石形,小淀粉体呈圆球形;(2)内胚乳细胞的生长呈“S”型曲线,花后16～24 d是细胞生长最快的时期;(3)大淀粉体的增殖和生长主要在花后4～16 d进行,小淀粉体在花后16 d大量出现并快速增殖,至胚乳发育后期小淀粉体数量占内胚乳细胞淀粉体总量的90％以上;(4)内胚乳细胞核在颖果发育中、后期发生衰亡,淀粉体在核衰亡过程中仍然能增殖和生长.     

干旱胁迫下小麦颖果内源激素的变化及其与胚乳发育的关系

为探明干旱胁迫下小麦颖果内源激素与胚乳发育的关系,以小麦品种扬麦16为材料,在幼苗返青至颖果成熟阶段进行干旱处理,采用树脂切片及显微摄影等技术观察小麦颖果和胚乳细胞发育的形态结构特征,并通过酶联免疫吸附法测定颖果生长素(IAA)、脱落酸(ABA)、赤霉素(GA_3、GA_4)、玉米素核苷(ZR)、二氢玉米素核苷(DHZR)的含量。结果显示,干旱胁迫缩短了小麦颖果发育进程,促进颖果早衰,导致颖果发育不良;显著降低了籽粒千粒重、可溶性糖含量和总淀粉含量,提高了蛋白质含量;促进颖果发育早期胚乳细胞分裂和淀粉积累,抑制了发育后期胚乳细胞的分裂、体积扩大和淀粉体充实,并提高了颖果整个发育过程中胚乳蛋白体的积累;提高了颖果发育前期IAA、GA_3、GA_4、ZR和DHZR的含量,ABA含量在整个发育时期均较高。说明干旱胁迫能通过影响颖果内源激素的积累来调控胚乳发育。     

小麦穗部和颖果维管束系统的发育解剖学研究

为了研究小麦穗部和颖果维管束的特性,以中筋小麦品种扬麦16为材料,通过光学、电子和荧光显微镜等观察和分析了穗部和颖果中维管束的结构、数目、发育及其相互关系.结果表明:(1)在花后8～28 d内,小穗轴中央维管束被TTC染色很红,随后变浅;从花后第4 d起,小穗轴中央维管束对I2-KI着色,呈色持续至花后24 d,其后不呈色.(2)花后1～6 d,颖果腹部维管束未完全分化成型,花后12 d,较完善的输导组织已发育形成,花后15～30 d,腹部维管束发育成熟并发挥物质运输作用,花后36 d,维管束输导功能已基本丧失.(3)花后19 d,颖果维管束中筛分子有典型的伴胞,并形成筛分子-伴胞复合结构,周围有大型的薄壁细胞,伴胞或薄壁细胞的体积比筛管分子大,细胞质浓,线粒体密度大;花后32 d,颖果韧皮薄壁细胞外形不规则,许多细胞壁突出或内陷,大部分韧皮薄壁细胞核解体,线粒体数目变少.(4)穗轴节片维管束排列为内、外两环,外环的维管束较小,位于茎的边缘,内环的维管束较大,周围为基本组织所包围.(5)穗轴节片大维管束数目从下而上逐减,上部和下部节片维管束下降幅度较少,而中部下降较多,但小维管束数目变化与此相反.(6)小穗轴基部维管束为12～13条,每小穗内的第1小花基部维管束为6～7条,第2小花中为5～6条,第3、4小花中为3～4条,第5小花以上各花中均为3条.     

不同类型专用小麦品种胚乳发育的比较研究

为揭示不同类型专用小麦品种胚乳发育的差异,以强筋小麦皖麦38和弱筋小麦扬麦9号为材料,比较了两者胚乳细胞游离核分裂状况、细胞数目的增殖变化、细胞体积的变化、糊粉层发育、淀粉体和蛋白质体发育等内容,主要结果如下：（1）皖麦38游离核有丝分裂所占比例较大,扬麦9号无丝分裂所占比例较大,且受温度影响较大;（2）胚乳细胞增殖均呈"S"型曲线变化,皖麦38胚乳细胞增殖较快,胚乳体积较大,最终粒重较高;（3）皖麦38胚乳细胞中大淀粉体数目多于扬麦9号,而小淀粉体数目表现却相反;（4）皖麦38与扬麦9号相比较,糊粉层出现时间较晚,而且糊粉层细胞壁较厚,被甲苯胺蓝染色更浓;（5）成熟籽粒中淀粉粒形状有饼形、椭圆形和近圆球形三种。皖麦38胚乳中饼形和椭圆形淀粉粒较多,且相互结合较紧密;扬麦9号饼形和近球形淀粉粒较多,胚乳结构疏松。     

Structural and Histochemical Characterization of Developing Rice Caryopsis

The development of pericarp, seed coat, starchy endosperm and aleurone of the rice caryopsis was investigated, histochemically and structurally, from the time of flowering to maturity. The results showed that during its growth, the maximum length of the caryopsis was attained first, followed by width and then thickness. Histochemical examination of the caryopsis showed that starch was mainly accumulated in the endosperm, but the endosperm showed no metabolic activity, while embryo and pericarp contained a few starch grains, and embryo and aleurone were strongly active. Aleuronic cells contained many aleurone grains and spherosomes, and aleurone in the dorsal region developed earlier and contained more layers of cells. Amyloplasts in endosperm contained many starch granules and were spherical at early stages but polyhedric at late stages. The protein bodies appeared later than amyloplasts, and the number of protein bodies in subaleurone was greater than those in the starchy endosperm. The white-belly portion of endosperm might be relative to the status of amyloplast development.     

Evaluation of Tissue Dissociation of Durum Wheat Grain (Triticum durum Desf.) Generated by the Milling Process

The three major botanical components (starchy endosperm, aleurone layer and pericarp) of eight durum wheat samples exhibited significantly different compositions and concentrations in phenolic acids. The starchy endosperm, the aleurone layer and the pericarp were respectively characterised by a low content in ferulic acid (FA), a high content intrans -sinapic acid (t -SA), and a high content in ferulic acid dehydrodimers (DHD). These three chemical markers can be exploited to differentiate the three grain botanical parts within milling fractions and to evaluate the milling efficiency, particularly the separation between bran and endosperm. The histological dissociation of the wheat grain generated by the milling process can be investigated further into details using the three phenolic acids markers. A separability index (S i) was proposed in order to quantify the ease of dissociation of endosperm from bran. Differences in S i values between wheat varieties grown under various agricultural conditions demonstrated the relevant variability of this character. The structural and molecular factors implied in the control of tissue dissociation are discussed in details.     

Spatial distribution of functional components in the starchy endosperm of wheat grains

The starchy endosperm of the mature wheat grain comprises three major cell types, namely sub-aleurone cells, prismatic cells and central cells, which differ in their contents of functional components: gluten proteins, starch, cell wall polysaccharides (dietary fibre) and lipids. Gradients are established during grain development but may be modified during grain maturation and are affected by plant nutrition, particularly nitrogen application, and environmental factors. Although the molecular controls of their formation are unknown, the high content of protein and low content of starch of sub-aleurone cells, compared to the other starchy endosperm cells types, may result from differences in developmental programming related to the cells having a separate origin (from anticlinal division of the aleurone cells). The gradients within the grain may be reflected in differences in the compositions of mill streams, particularly those streams enriched in the central and outer cells of the starchy endosperm, respectively, allowing the production of specialist flours for specific end uses.     

Endosperm Development in Autotetraploid Rice

By using the laser scanning confocal microscope and plastic （Leica 7022 historesin embedding kit） semi-thin sectioning technique, comparative studies on the development of endosperm were carried out between autotetraploid and diploid rices. About one third of the ovaries in the autotetraploid showed normal endosperm development as those in the diploid. In these ovaries, one of the polar nuclei would fuse with the sperm nucleus, and the primary endosperm nucleus formed and underwent the first division in 4 hours after pollination; the anticlinal wall began to grow centripetally between the free nuclei starting from the wall ingrowths of the embryo sac near the micropylar end, and some of the phragmoplasts formed transformed into periclinal walls. In addition, some of the cell wall situated in the middle of the endosperm appeared to originate from phragmoplasts, whereas others seemed to develop randomly without the obvious formation of phragmoplasts. Cellulose began to accumulate in the wall of aleurone cell layer at 6 days after pollination. The cellulose wall of the cells of the aleurone cell layer appeared to have completely formed within 7 to 8 days after pollination. On the other hand, about two thirds of the ovaries in the autotetraploid showed abnormality in endosperm development with various types, such as non-fertilization, abnormal fertilization, endosperm development-delay and non-synchronization in the development of cellulose wall of cells of the aleurone layer. These abnormalities usually resulted in decreased seed setting in autotetraploid rice.