Effects of inter-culture, arabinogalactan proteins, and hydrogen peroxide on the plant regeneration of wheat immature embryos

The regeneration rate of wheat immature embryo varies among genotypes, howbeit many elite agriculture wheat varieties have low regeneration rates. Optimization of tissue culture conditions and attempts of adding signal molecules are effective ways to increase plant regeneration rate. Inter-culture is one of ways that have not been investigated in plant tissue culture. Moreover, the use of arabinogalactan proteins （AGPs） and hydrogen peroxide （H202） have been reported to increase regeneration rate in a few plant species other than wheat. The current research pioneeringly uses inter-culture of immature embryos of different wheat genotypes, and also investigates impacts of AGP and H2O2 on the induction of embryogenic calli and plant regeneration. As a result, high-frequency regeneration wheat cultivars Kenong 199 （KN 199） and Xinchun 9 （XC9）, together with low-frequency regeneration wheat line Chinese Spring （CS）, presented striking increase in the induction of embryogenic calli and plant regeneration rate of CS through inter-culture strategy, up to 52.19 and 67.98%, respectively. Adding 50 to 200 mg L-1 AGP or 0.005 to 0.01‰ H2O2 to the callus induction medium, enhanced growth of embryogenic calli and plant regeneration rate in quite a few wheat genotypes. At 50 mg L-1 AGP application level in callus induction medium plant regeneration rates of 8.49,409.06 and 283.16% were achieved for Jimai 22 （JM22）, Jingdong 18 （JD18） and Yangmai 18 （YM18）, respectively; whereas at 100 mg L-1 AGP level, CS （105.44%）, Chuannong 16 （CN16） （80.60%） and Ningchun 4 （NC4） （62.87%） acted the best. Moreover CS （79.05%）, JM22 （7.55%）, CN16 （101.87%）, YM18 （365.56%）, Yangmai 20 （YM20） （10.48%）, and CB301 （187.40%） were more responsive to 0.005 %o of H2O2, and NC4 （35.37%） obtained the highest shoot regeneration rates at 0.01%o of H2O2. Overall, these two methods, inter-culture and AGP （or H2O2） application, can be further applied to wheat transgenic research.

Identification and functional analysis of arabinogalactan protein expressed in pear pollen tubes

Arabinogalactan proteins(AGPs)are widely distributed in the plant kingdom and play a vital role during the process of plant sexual reproduction.In this study,we performed a comprehensive identification of the PbrAGPs expressed in pear pollen and further explored their influences on pollen tube growth.Among the 187 PbrAGPs that were found to be expressed in pear pollen tubes,38 PbrAGPs were specifically expressed in pollen according to the RNA-seq data.The PbrAGPs were divided into two groups of highly expressed and specifically expressed in pear pollen.We further tested their expression patterns using RT-PCR and RT-qPCR.Most of the PbrAGPs were expressed in multiple tissues and their expression levels were consistent with reads per kilobase per million map reads(RPKM)values during pollen tube growth,implying that PbrAGPs might be involved in the regulation of pear pollen tube growth.We also constructed phylogenetic trees to identify the functional genes in pear pollen tube growth.Therefore,19 PbrAGPs(PbrAGP1 to PbrAGP19)were selected to test their influences on pollen tube growth.Recombinant proteins of the 19 PbrAGP-His were purified and used to treat pear pollen,and 11 of the PbrAGP-His recombinant proteins could promote pear pollen tube growth.Additionally,pollen tube growth was inhibited when the expression levels of PbrAGP1 and PbrAGP5 were knocked down using an antisense oligonucleotide assay.PbrAGP1 and PbrAGP5 were localized in the plasma membrane and might not alter the distribution of pectin in the pollen tube.In summary,this study identified the PbrAGPs expressed in pear pollen and lays the foundation for further exploring their functions in pollen tube growth.

DEAP1 encodes a fasciclin-like arabinogalactan protein required for male fertility in rice

Arabinogalactan proteins(AGPs)are widely distributed in plant cells.Fasciclin-like AGPs(FLAs)belong to a subclass of AGPs that play important roles in plant growth and development.However,little is known about the biological functions of rice FLA.Herein,we report the identification of a male-sterile mutant of DEFECTIVE EXINE AND APERTURE PATTERNING1(DEAP1)in rice.The deap1 mutant anthers produced aberrant pollen grains with defective exine formation and a flattened aperture annulus and exhibited slightly delayed tapetum degradation.DEAP1 encodes a plasma membrane-associated member of groupⅢplant FLAs and is specifically and temporally expressed in reproductive cells and the tapetum layer during male development.Gene expression studies revealed reduced transcript accumulation of genes related to exine formation,aperture patterning,and tapetum development in deap1 mutants.Moreover,DEAP1 may interact with two rice D6 PROTEIN KINASE-LIKE3 s(OsD6PKL3s),homologs of a known Arabidopsis aperture protein,to affect rice pollen aperture development.Our findings suggested that DEAP1 is involved in male reproductive development and may affect exine formation and aperture patterning,thereby providing new insights into the molecular functions of plant FLAs in male fertility.

Immunolocalization of Arabinogalactan Proteins and Pectins in Floral Buds of Cucumber （Cucumis sativus L.） During Sex Determination

Abstract: Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber (Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207 (recognizes AGP epitopes); JIM 8 (recognizes a subset of AGP epitopes); and JIM 5 and JIM 7 (epitopes of pectins esterified to various degrees). In the stem apex meristem (SAM) of the cucumber, epitopes of MAC 207, JIM 7, and JIM 5 were localized in the cells from second to third peripheral layers when the sex organ primodium began to differentiate; epitopes of MAC 207 and JIM 5 were also detected in the ragged edge cells. A very dense labeling signal with MAC 207 was observed in the carpel and pistil primodium. The AGP epitopes recognized by JIM 8 were localized in the anther of the male flower and the anther-like portion of the stagnant stamen of the female flower. This suggests that the AGPs and pectins in the SAM of the cucumber are closely associated with the differentiation of the SAM, from meristematic cells to floral primodium. The subset of AGPs recognized by JIM 8 may play an important role in stamen formation.

Efficient Plant Regeneration with Arabinogalactan-Proteins on Various Ploidy Levels of Cereals

To determine the most effective dose of arabinogalactan-protein （AGP） in regeneration medium, mature embryos of genotypes in three different ploidy levels （Triticum aestivum L. cv. Ikizce-96, Triticum durum Desf. cv. Mirzabey and Hordeum vulgare L. cv. Tokak） were used to establish an efficient plant regeneration system for cereals. Percentage of callus production, capacity of regeneration were calculated, and also culture effect, root, stem, and total plant length of regenerant plants were observed in six different regeneration media （MS control, MS＋2, 5, 7, 10, 12 mg L-1 AGP） in these three different genotypes. According to the results, the highest amount of callus production was found in Ikizce-96 as 93.75% using 5 mg L-1 dicamba and 1 mg L-1 kinetin in induction medium. However, the most improved callus was observed in diploid barley Tokak as 179.95 mg in weight and 6.18 mm in diameter, respectively. The highest regeneration capacity was observed in the dose of 5 mg L-1 AGP in MS of all the genotypes and hexaploid wheat Ikizce-96 gave the best results with the highest regeneration capacity and culture effects （94.86 and 92.5%） in the same dose of AGE These results indicated that effective dose of AGP in regeneration medium increase plant regeneration in calli derived from cereal mature embryos.

果胶酶对灵武长枣果实发育中阿拉伯半乳糖蛋白分布的影响

以3种不同浓度的果胶酶处理灵武长枣(Ziziphus jujuba‘Lingwu Changzao’)4个不同发育时期的果实,采用免疫细胞化学技术,在细胞水平上对果实阿拉伯半乳糖蛋白(AGPs)表位进行原位分析,探讨果胶酶对其不同发育时期的果实中AGPs分布的影响,为明确果胶酶对果实成熟软化的影响提供解剖学依据。结果表明:JIM13、JIM8和MAC204抗体所标记的抗原荧光强弱在各时期果实的不同组织存在一定的差异。当果肉组织用较低浓度的果胶酶0.028 U·mL^(-1)(E1)处理,果皮组织结构没有明显的变化,果皮及内部薄壁细胞细胞壁表面和细胞间隙中的AGPs抗原表位减少;0.056 U·mL^(-1)(E2)和0.084 U·mL^(-1)(E3)浓度果胶酶处理的果实,果实组织细胞壁解体程度增加,果皮及内部薄壁细胞细胞壁中AGPs抗原表位检测量逐渐降低,果胶酶浓度的增加导致AGPs在果实所有表位排列的更大影响和较低的荧光信号。经果胶酶最高浓度0.084 U·mL^(-1)(E3)处理后,Calcofluor White染色显示细胞壁区域荧光也不同程度减弱或降低,AGPs分布的紊乱和抗原表位的缺失与细胞中纤维素组装的变化有关。比较分析表明,不同发育时期的果实AGPs碳水化合物的分布不同,这与组织结构的变化有关;果胶酶作用下AGPs聚糖链的缺失导致细胞壁组分之间的相关性建立和细胞壁结构的重塑受阻,诱导了整个细胞壁结构的改变,影响了果实的成熟。

巨桉FLA基因家族成员的鉴定与分析

【目的】含有束状蛋白(fasiclin)结构域的类成束状阿拉伯半乳糖蛋白(fasciclin-like arabinogalactan proteins,FLAs)在巨桉细胞壁的次生生长和合成中起着重要作用。为了获得更多有价值的信息并探索其功能,对巨桉FLA基因家族的全基因组进行分析。【方法】以巨桉全基因组数据和转录组数据为基础,通过Expasy、MEME、MEGA7.0、TBtools等生物信息学工具对EgrFLAs基因家族的核酸序列的特征、基因结构、蛋白质理化性质、蛋白保守结构域、家族系统进化关系和表达量等进行分析和预测。【结果】巨桉共有21个EgrFLAs基因,其中包括3个新发现的EgrFLAs基因,这些基因不均匀地分布在巨桉11条染色体中的9条上。所有鉴定到的EgrFLAs基因均含有束状蛋白结构域,聚为4个群组,其中D组包含最多的基因成员。在EgrFLAs基因启动子中发现了大量的顺式元件,包括发育相关、水杨酸、茉莉酸甲酯等相关响应元件。不同组织的表达谱分析表明除EgrFLA1、EgrFLA2和EgrFLA3外,聚在A组的EgrFLA5和EgrFLA7也参与了次生生长。在巨桉不定根形成过程中,EgrFLA1和EgrFLA3通过表达量逐渐增加的方式参与诱导不定根形成。经SA和MeJA处理后,EgrFLA1和EgrFLA2的表达量发生显著变化,表明EgrFLA1和EgrFLA2在巨桉的发育和胁迫反应中可能具有多种作用。【结论】在巨桉中鉴定FLA基因21个,包括新发现的3个基因。在EgrFLAs基因启动子中发现了大量发育和激素相关的顺式元件。EgrFLA1、EgrFLA2、EgrFLA3、EgrFLA5和EgrFLA7作为巨桉次生生长发育中的候选基因,将为进一步的巨桉分子育种提供宝贵的资源。

桃金娘果实多糖的构造研究(I)

对新鲜和自然干燥后桃金娘的果实中的水溶性多糖进行了分离和化学构造初步研究。结果表明桃金娘多糖是木聚糖、阿拉伯半乳聚糖(AG)或者阿拉伯半乳聚糖糖蛋白(AGP)和果胶等构成的混合多糖。通过鲜果与干果实验数据的比较,其水溶性多糖的构成以及各组分含量均有明显变化。水溶性多糖分离纯化所得到的H3和H4组分的构成单糖分析表明,它们可能是含有较多毛发区域(鼠李半乳糖醛酸聚糖为主链)的果胶多糖。

烟草柱头和花柱中阿拉伯半乳糖蛋白的定位

通过Western印迹法、免疫组织化学和超微细胞化学等技术,研究了烟草柱头和花柱中阿拉伯半乳糖蛋白(arabinogalactan-proteins,AGPs)的分布。结果表明烟草柱头和花柱组织中含有大量的AGPs,主要分布于柱头表皮细胞的细胞质和分泌层细胞的胞外基质中,且授粉前后AGPs的分布情况差异不明显;而花柱中的AGPs主要分布于表皮细胞的外层细胞壁、维管组织周围细胞的细胞质及引导组织的胞外基质中;花粉管通过后,引导组织胞外基质中AGPs减少,而花粉管细胞质和花粉管壁中检测到大量AGPs。

香蕉胚性及非胚性悬浮培养的建立及其培养液中阿拉伯半乳糖蛋白含量的测定

测定了贡蕉花序外植体起始的胚性细胞悬浮系以及东莞大蕉薄片外植体起始的非胚性细胞悬浮系中,悬浮细胞分泌到培养基中的AGPs的含量。结果显示,香蕉细胞悬浮系在培养过程中分泌到培养基中的AGPs浓度与细胞系的发育状态相关,胚性细胞分泌的AGPs的含量高于非胚性细胞。该结果为今后研究AGPs与香蕉悬浮细胞胚性潜能的关系提供了可能性。

枸杞子阿拉伯半乳糖聚糖糖蛋白的微细构造研究(I)

对从枸杞子水溶性粗多糖中分离出来的一种主要阿拉伯半乳糖聚糖糖蛋白Cp-2-B的微细构造进行了研究.通过高碘酸氧化和缓和Smith分解,从分解产物中分离出一个分子量为1.8万的半乳糖聚糖 (Cp-2-B-SD)。 甲基化分析和GC-MS分析结果表明Cp-2-B-SD为典型的梳状构造:它的主链是由(1→3)结合的半乳糖残基组成,其中约30%的半乳糖残基带有取代基。另外,还通过蛋白质水解酶反应以及阿拉伯半乳糖聚糖糖蛋白(AGP)与β-galactosylYariv试剂的特有免疫反应等手段,探讨了Cp-2-B的微细构造特征。

βGlcY影响油菜小孢子胚胎发生起始分裂

以油菜小孢子为材料,研究不同浓度的βGlcY对小孢子胚胎发生起始分裂的影响,结果表明,βGlcY对油菜小孢子胚胎发育起始细胞分裂的抑制作用是浓度依赖的行为.βGlcY浓度为50μmol/L时,小孢子的分裂完全受到抑制.细胞学研究发现,βGlcY与AGPs结合后影响了小孢子的活性,推测βGlcY可能诱导了小孢子的程序化死亡.

阿拉伯半乳糖蛋白在被子植物有性生殖中的作用

阿拉伯半乳糖蛋白(arabinogalactan-proteins,AGPs)是一类主要分布在细胞表面和胞外基质中的糖蛋白。它们在植物的雄性器官(花粉、花粉管、精细胞)、雌性器官(柱头、花柱、子房)和胚胎(合子胚和体细胞胚)等组织和细胞中均有大量的表达。大量研究表明AGPs 在被子植物有性生殖过程中起着非常重要的作用,既可能参与花粉管粘附、营养、传导或提供信号的作用,也可能参与受精过程中配子识别和受精后胚胎的发育与分化等过程。该文就其分子结构、特性以及在植物有性生殖过程中各种器官和组织内的表达和功能研究进展做了较为全面的概述。

灵武长枣果实阿拉伯半乳糖蛋白免疫荧光定位

目的:为探讨灵武长枣果实AGPs糖蛋白在果实发育过程中的分布特征和动态变化规律。方法:以4个时期灵武长枣果实为实验材料,通过免疫组织化学方法,研究了不同发育时期果实阿拉伯半乳糖蛋白AGPs的分布,为进一步研究灵武长枣果实AGPs在品质调控方面的功能奠定基础。结果:(1)JIM13和JIM8抗体所识别的抗原在4个时期果实的外果皮及相邻的内部数层排列紧密的中果皮小细胞细胞壁和细胞内部均有分布。(2)除了空腔之外,内部的中果皮大型卵圆形薄壁细胞的细胞壁和细胞内在膨大前期均有JIM13和JIM8抗体所识别的抗原分布,而快速膨大期、着色期和完熟期果实JIM13和JIM8抗体所识别的抗原均主要分布于细胞壁上,大部分细胞内部无分布;随着果实发育成熟,空腔数量增多,中果皮薄壁细胞间隙拉大排列更加松散,出现细胞破裂,JIM13和JIM8抗体所识别的抗原分布逐渐减少。(3)4个时期果实维管束中维管束鞘、木质部、韧皮部、形成层所有细胞的细胞壁和细胞内部都分布有JIM13和JIM8抗体所识别的抗原,维管束数量和大小随果实发育及体积的进一步增大逐渐减少,JIM13和JIM8抗体所识别的抗原分布逐渐减少。结论:JIM13和JIM8为灵武长枣果实免疫组织化学的良好抗体,各时期果实的不同组织JIM13和JIM8抗体所识别的抗原荧光强弱存在一定的差异;AGPs参与了灵武长枣果实维管束发育过程的形态建成、细胞的分裂和体积的增大,为果实发育提供营养支持及保护。

阿拉伯半乳糖蛋白在高等植物中的分布及功能

阿拉伯半乳糖蛋白(arabinogalactan-proteins,AGPs)是一类含有丰富羟脯氨酸或脯氨酸的结构复杂的糖蛋白,其广泛分布在细胞壁、原生质体和质膜上。在植物的根部、花粉管、花粉以及柱头、花柱等组织中均有表达,AGPs在被子植物营养生长和生殖发育均起着非常重要的作用,可能参与了根形态建成与发育,根系与土壤微生物互相作用,并影响花粉管的生长、花粉粒的成熟和柱头上萌发等。现就其分子结构、分类以及主要研究方法,对植物根组织以及花粉、花粉管、柱头部位AGPs的分布和可能功能进行了综述。

A suppressed gene in integument cells of a fiberless seed mutant in upland cotton

A fiberless seed mutant (fl) was identified in a commercial cotton ( Gossypium hirsutum L.) variety Xu Zhou 142 (FL). This phenotype is associated with lack of fiber cell initiation in the outer integument of the ovule, as was characterized by analysis of genes related to fiber differentiation and development. Two genes, fl E6 and FL E6, were cloned from fl integument cells and FL fiber or integument cells, respectively. Compared with FL E6, fl E6 showed a dramatic change in nucleotide sequence: (1) FL E6 contained a tandem repetitive sequence in which GGCTCA (Gly Ser) is repeated five times between the 82nd and the 93rd codon from the first ATG codon, while in fl E6 the same sequence is repeated four times; (2) The fl E6 gene encodes a polypeptide of 241 amino acids but lacks two codons between the 90th and 93rd codon and three between the 171st and 174th relative to FL E6; (3) There are also 12 nucleotide substitutions which would result in 7 amino acid differences between fl E6 and FL E6. Analysis of RT PCR and Northern Blot showed that expression of the fl E6 gene is suppressed in the fl integument cells, but highly expressed in FL fiber cells. The difference between fl E6 and FL E6 may be associated with lower expression of fl E6 in the fl integument cells. Searches of protein databases with the FL E6 gene sequence showed similarity to the protein backbones of two arabinogalactan proteins (AGPs), one from the filtrate of suspension cultured cells of Pyrus communis (AGPPc2) and the other from Nicotiana alata (AGPNa2). Although the function of the FL E6 protein in differentiation and development of cotton fiber cells is not known, the data indicate that the mutation of fl E6 gene from FL E6 gene may inhibit the fiber cell initiation from epidermal cells of the outer integument of the ovule.

枸杞子阿拉伯半乳糖聚糖糖蛋白的微细构造研究(Ⅱ)

按照Taylor法(还原剂为NaBD4)将从枸杞子中分离出来的阿拉伯半乳糖聚糖糖蛋白Cp-2-B中含有的半乳糖醛酸还原成重氢标记的半乳糖,对还原前后的样品进行甲基化处理后供GC-MS。 结果发现Cp-2-B分子中存在着非还原末端,(1→3)- 结合以及(1→4)- 结合等3种半乳糖醛酸残基。核磁共振分析也证实了这一结论。另外,从Cp-2-B与果胶粗酶以及果胶内切酶的反应结果可以推测出,半乳糖醛酸残基以分散状态或者短链形式存在,分子中不存在类似果胶物质那样的、由连续的(1→4)结合的半乳糖醛酸组成的Smooth领域。

兴安落叶松温水提取液生产单细胞蛋白的研究

对兴安落叶松温水提取液的水解及其用于生产单细胞蛋白的可能性进行了研究。未经水解的提取液主要含有阿拉伯-半乳聚糖,难以被试验菌种利用;水解后添加必要的氮、磷等营养物质,被试菌种可利用其中的单糖生产单细胞蛋白。拟青霉能利用阿拉伯糖、半乳糖等多种糖类,菌体产率最高,占总糖的51.9％;热带假丝酵母、产朊假丝酵母部分或完全不能利用占总糖20％的阿拉伯糖,菌体产率分别为40.98％和36.00％。商品半纤维素酶对阿拉伯-半乳聚糖的降解活性不高,但自制的粗酶能较好地水解阿拉伯-半乳聚糖,水解率可达71.85％。

阿拉伯半乳糖蛋白对香蕉悬浮细胞胚性状态的影响

采用火箭琼脂糖凝胶电泳法测定了培养15 d后的香蕉悬浮细胞液体培养基中的阿拉伯半乳糖蛋白(ara-binogalactan-proteins,AGPs)的含量,发现具有体胚发生能力的贡蕉、大蕉和龙牙蕉过山香品种的胚性悬浮细胞(em-bryogenic suspension cells,ECS)培养基中的AGPs含量分别达到36、48、56 mg.L-1,而在不能进行体胚诱导的香芽蕉威廉斯品种的非胚性悬浮细胞(non-embryogenic suspension cells,NECS)的培养基中检测不到AGPs,说明悬浮细胞分泌到培养基中的AGPs含量与香蕉悬浮细胞的胚性状态有一定的关系。在香蕉ECS的悬浮培养过程中,添加不同浓度的βGlcY试剂(β-glucosyl Yariv)到悬浮培养基中可引起胚性细胞与非胚性细胞(ECS/NECS)的比值下降;在香蕉的ECS体胚诱导过程中,添加不同浓度βGlcY试剂到体胚诱导培养基中,引起体胚发生频率下降。这些结果表明,AGPs在香蕉悬浮细胞的胚性保持过程中起到重要的作用,并且具有剂量效应。

化学杀雄剂SQ-1和阿拉伯葡聚糖蛋白对小麦品种间杂交及远缘杂交成胚率的影响

【目的】探究化学杀雄剂SQ-1对小麦品种间、小麦与近缘植物间、小麦与远缘植物间杂交成胚率的影响,以及阿拉伯葡聚糖蛋白对小麦与玉米杂交成胚率和得苗率的影响。研究结果对于合理选用小麦杂交方式,提高小麦杂交结实率和利用玉米诱导小麦单倍体植株的效率具有重要意义。【方法】通过在小麦拔节期喷施化学杀雄剂SQ-1,开花期分别授以小麦花粉和远缘植物(黑麦、玉米)花粉,并在小麦授玉米花粉后的处理液中加入不同浓度的阿拉伯葡聚糖蛋白(arabinogalactan proteins,AGP),对小麦与玉米杂交后产生的幼胚进行离体拯救培养,统计授粉小花数、接种幼胚数、膨大颖果数、结实粒数、萌发单倍体幼胚数和单倍体植株数,计算结实率、颖果膨大率、成胚率、萌发率和成苗率并对所得数据进行差异显著性分析,结合细胞学观察结果,研究SQ-1对小麦品种间杂交及远缘杂交结实性的影响,以及AGP对小麦单倍体胚诱导率的影响。【结果】在不同小麦品种间杂交中SQ-1处理结实率19.8%—83.3%,人工去雄的结实率为69.4%—93.0%,SQ-1对不同品种的影响不同,Fielder对SQ-1的反应比较敏感;在中国春与兰州黑麦杂交中,SQ-1处理的结实率为65.5%,人工去雄处理的结实率为78.8%,两种处理方式产生的F1杂种的染色体数均为28条;在不同小麦品种与玉米品种郑单58杂交中,SQ-1处理小麦单倍体胚的成胚率为1.11%—1.41%,人工去雄小麦单倍体胚的成胚率为2.38%—14.29%;在小麦与玉米杂交后的处理液中添加0.5—2.0 g·L-1 AGP一定程度地提高了小麦单倍体胚获得率和成苗率。另外,在玉米花粉诱导的单倍体胚离体培养过程中,发现13.07%的胚发育出了2—6株苗;显微镜观察发现,玉米花粉诱导后18 d左右小麦单倍体胚上出现了多个突起,这些突起在离体培养条件下进一步发育为形态健全的小植株,其染色体数目均为21条。【结论】化学杀雄剂SQ-1减低了小麦品种间杂交及小麦与黑麦、玉米间杂交的成胚率,AGP提高了小麦与玉米间杂交单倍体成胚率和成苗率。