小麦A，B和D基因组同源DNA序列在进化过程中的演变研究

选取已定位的大麦1H染色体的STS标记NWG913为引物,在普通小麦（Tritium aestivum L.）及其4个可能的起源种乌拉尔图小麦（T.urartu T.)、栽培一粒小麦（T.monococcum.L)栽培二粒小麦（T.dicoccum S.）、方穗山羊草（Ae.squarrosa L.)上特异性扩增。扩增产物克隆测序后对其进行序列分析,由序列差异的程度来确定这几个物种之间的亲缘关系。实验结果表明,普通小麦（Tritium aestivum L.)的A基因组此段序列与乌拉尔图小麦（T.urartu T.)、栽培一粒小麦（T.monococcum L.)、栽培二粒小麦（T.dicoccum S.)A基因组此段序列完全相同;普通小麦的D基因组此段序列与方穗山羊草（Ae.squarrosa L.)也完全相同;普通小麦的B基因组此段序列和栽培二粒小麦B基因组此段序列有0．61％的差异。研究结果一方面对现有的普通小麦A、B、D基因组起源和进化理论给予了分子水平上的证明,同时也揭示了同一物种不同的基因组化进化速度存在差异。     

小麦进化过程中叶片气孔和光合特征演变趋势

根据小麦属内种间的进化关系选取21种小麦品种为实验材料,研究了小麦进化过程中气孔特征和光合特征的演变趋势。结果表明,无论是A,B,D染色体组还是A,G染色体组,气孔长度、宽度、周长、面积均随倍性水平的升高而呈增大趋势,而气孔指数无显著变化;A,B,D染色体组气孔密度随倍性升高呈减少趋势。二倍体小麦的Pn值最高,六倍体小麦的Fv/Fm值较高,二倍体小麦叶片叶绿素含量显著高于四倍体和六倍体小麦。不同倍性小麦的净光合速率与气孔导度间均存在极显著相关关系,表明气孔传导力是小麦光合能力的主要限制因素之一。气孔导度与单一气孔特征之间无显著相关关系。A,B,D染色体组不同倍性小麦叶片气孔密度差异显著,其大小顺序为二倍体〉四倍体〉六倍体;A,B,D染色体组不同倍体小麦叶片的气孔长度、宽度、周长、面积差异显著,顺序均为六倍体〉四倍体〉二倍体,气孔密度降低可能是A,B,D染色体组六倍体小麦光合能力降低的原因。随着倍性的升高,小麦的抵抗光抑制能力越强,因此光化学能转换效率可能不是小麦进化过程中光合能力变化的原因。A,B,D染色体组中二倍体的叶绿素含量显著大于四倍体和六倍体,而A,G染色体组中倍体间叶绿素含量差异不显著,说明叶绿素的降低可能是A,B,D染色体组六倍体光合能力降低的原因之一。     

水分胁迫对小麦叶片光合作用的影响及其与抗旱性的关系

在水分胁迫初期,两个小麦品种叶片光合速率,气孔导度和细胞间隙CO_2浓度降低,气孔限制值增加,光合速率的降低主要是气孔因素的限制。中度到严重水分胁迫使叶片光合速率、气孔导度和气孔限制值降低,细胞间隙CO_2浓度明显增加,且叶圆片放氧能力,叶绿体Hill反应、叶绿素荧光强度和表观量子产额降低,此时光合速率的降低主要是叶肉细胞光合活性的下降引起的。抗旱性弱的郑引一号叶肉细胞光合活性比抗旱性强的丰抗13更容易受到水分胁迫的影响。     

砷对小麦生长和光合作用特性的影响

研究了水培条件下砷对小麦根系和地上部分生长速率、光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)和蒸腾速率(Tr)的影响.结果表明,随着营养液中砷浓度的提高,小麦根长生长量和地上部分生长量较对照减少;鲜重和砷的浓度呈显著负相关.在0～90mg/L 砷处理内,Pn、Gs、Tr都随砷的浓度的提高而降低;Ci呈先降低,后升高的变化.在As≤30 mg/L时Ci逐渐降低,气孔限制值Ls升高,使Pn下降,造成气孔性限制;而As>30 mg/L时,Ci升高,气孔限制值Ls下降,Pn降低,造成了非气孔性限制.叶片水分利用效率WUE和气孔限制值Ls在As≤30 mg/L时变化一致,都有所升高;但是当As>60 mg/L时,小麦趋于死亡,水分利用率降低.叶绿素含量在0～10mg/L As处理内,差异不显著,在较高砷浓度(As>30 mg/L)时叶绿素显著下降.这说明较低浓度的砷不会抑制小麦叶绿素的合成.砷主要是毒害小麦根系生长,造成植株体光合作用的气孔性限制和非气孔性限制出现,最终影响小麦的生长和发育.     

用ITS序列确定小麦B基因组的可能供体间的关系

对小麦B基因组的可能供体山羊草属Aegilops sect．Sitopsis的5个种的核糖体DNA的内部转 录区(ITS)进行了PCR扩增和克隆,井测定ITSl和ITS2的序列,用ITSl+ITS2的序列重建了 Aegilops sect．Sitopsis中5个种的系统发育关系。结果表明,斯卑尔脱山羊草Ae．speltoides是sect． Sitopsis中特殊的一个种,它与该组其余4种间的平均遗传距离是后者彼此间平均遗传距离的3倍,Ae． speltoides与同组其余4个种的分离要比后者相互间的分离早得多;在拟斯卑尔脱组Sect．Sitopsis的5 个种中,长柱山羊草Ae．longissima与沙融山羊草Ae．sharonensis的关系最近。ITS序列可以进一步用来作为确定B基因组起源的分子标记。     

盐胁迫抑制槲栎2变种光合作用的机理研究

以不同浓度NaCl处理北京槲栎和锐齿槲栎的2年生幼苗,研究盐胁迫条件对其生长、光合作用及叶绿素荧光特性的影响。结果表明,(1)随着盐浓度的增加,总叶面积、植株鲜重和干重、叶片净光合速率(Pn)、气孔导度(Gs)、气孔限制值(Ls)、叶绿素含量(Chl)和光系统Ⅱ光合电子传递量子效率(ФPSⅡ)均显著降低,而光系统Ⅱ的初始荧光(F0)和光化学效率(Fv/Fm)无显著变化。(2)北京槲栎生长受盐胁迫的干扰小于锐齿槲栎,其净光合速率下降幅度相对较小,光合结构受胁迫的程度小于锐齿槲栎。     

等渗NaCl、干旱胁迫对番茄幼苗光合特性及叶绿体超微结构的影响

为分析等渗盐分和水分胁迫对番茄叶片光合功能的影响,选用耐盐性不同的4种基因型番茄(小果型的辽园红玛瑙、野生醋栗番茄、大果型的金田粉冠、超402)进行等渗的140 mmol·L^-1NaCl和15%PEG6000模拟盐分和水分胁迫.结果表明: 在光合特性方面,处理12 d后,两种胁迫导致4种不同基因型番茄的叶绿素含量(叶绿素a、叶绿素b、叶绿素总量)、光合速率、气孔导度、胞间CO₂浓度、蒸腾效率降低,气孔限制值升高.两种胁迫导致的光合下降由气孔性因素所致.等渗的盐分胁迫对番茄的光合系统损伤大于水分胁迫,这是因为盐分胁迫除了渗透胁迫还会导致离子伤害.4种不同基因型番茄中,耐盐型的辽园红玛瑙具有高光合特性,金田粉冠光合效率最差.在叶绿体超微结构方面,两种胁迫会造成番茄叶片的气孔密度增加,气孔张开率降低,叶绿体长度增加,宽度变小,长宽比增大,叶绿体内基粒数减少,嗜锇颗粒数增多.盐分胁迫下两种番茄叶绿体结构的影响大于水分胁迫,耐盐种醋栗番茄气孔变化小于盐敏感品种金田粉冠.     

不同小麦进化材料生育后期光合特性和产量

以二倍体野生一粒小麦(Triticum boeoticum)、栽培一粒小麦(T. monococcum)、节节麦(Aegilops tauschii)和黑麦(Secale cereale)、四倍体野生二粒小麦(T. dicoccoides)、栽培二粒小麦(T. dicoccum)、硬粒小麦(T. durum)、六倍体普通小麦(T. aestivum)‘扬麦9号’和‘扬麦158’及八倍体小黑麦(Triticale)为材料,采用盆栽试验研究了不同小麦进化材料生育后期旗叶光合特性的演变及产量的差异。结果表明,与六倍体普通小麦和八倍体小黑麦相比,二倍体和四倍体材料在开花前具有较高的光合速率(P_n)、气孔导度(G_s)、最大光能转换效率(F_v/F_m)和实际光化学效率(Φ_PSⅡ)。开花以后,二倍体和四倍体材料受非气孔因素的影响,光合能力下降较快;除黑麦外,旗叶光合速率在开花10 d后都低于普通小麦和小黑麦,胞间CO₂浓度(C_i)迅速增加,F_v/F_m、Φ_PSⅡ和叶绿素含量快速下降。二倍体和四倍体材料开花前单株总叶面积和旗叶叶面积较大,花后下降迅速,功能期短;单株穗数也较多,但穗粒数、千粒重、产量和收获指数却显著低于普通小麦。因此,小麦长期进化过程中,普通小麦花后较高的光合能力及较长的光合持续期是提高千粒重,进而提高产量的重要生理基础。     

纳米SiO2对髯毛箬竹叶片光合特征参数日变化的影响

采用叶面喷施的方法,研究了0(CK)、150、300和450 mg.L-1纳米SiO2对髯毛箬竹(Indocalamus barbatusMcClure)叶片净光合速率、气孔导度、蒸腾速率、胞间CO2浓度、气孔限制值和水分利用效率等光合特征参数日变化(7:00至16:00)的影响。结果表明:对照组髯毛箬竹叶片的净光合速率、气孔导度、蒸腾速率和气孔限制值的日变化曲线均为"双峰"型,有明显的"午休"现象;胞间CO2浓度的日变化曲线则为"W"型;水分利用效率的日变化曲线为"U"型。与对照相比,喷施150、300和450 mg.L-1纳米SiO2均能不同程度提高髯毛箬竹叶片的净光合速率、气孔导度和蒸腾速率,减小气孔限制值及胞间CO2浓度的变化幅度,提高"午休"时的水分利用效率,缓解光合作用的"午休"现象;其中,300 mg.L-1纳米SiO2处理组的效果最佳,在出现最大峰值的10:00时300 mg.L-1纳米SiO2处理组的净光合速率、气孔导度和蒸腾速率分别比对照提高了13.98%、71.21%和49.09%,且均与对照差异显著(P<0.05)。综合分析结果说明:髯毛箬竹叶片光合能力的提高是气孔和非气孔限制因素共同作用的结果,而喷施适量的纳米SiO2能够在一定程度上改善非气孔限制因素,从而增强髯毛箬竹叶片的光合能力。     

NaCl胁迫对大米草光合·蒸腾等生理特性的影响

研究了NaCl胁迫下大米草净光合速率(Pn)、蒸腾速率(Tr),叶片气孔导度(Gs)、细胞问CO_2浓度(Ci)、株高、叶长、叶宽、茎粗、叶绿素含量、气孔限制值和水分利用效率.结果表明:当NaCl浓度高于300mmol/L时,大米草Pn、Tr、Gs、株高、叶长以及叶绿素含量受到显著抑制(P＜0.05),叶宽及茎粗则无显著性差异.NaCl胁迫下,大米革光合速率的降低是气孔因素和非气孔因素综合导致的结果,Pn、Tr、Gs、株高以及叶绿素含量的降低可作为大米草受NaCl胁迫的症状,而WUE则保持在较高的水平,因此在防治大米草蔓延时,排水处理不是最佳选择.     

Differences in stomatal and photosynthetic characteristics of five diploidy wheat species

Five diploidy wheat species were chosen as research materials to determine the differences in stomatal and photosynthetic characteristics. The results of the study showed that species with S genomes (Ae. Sect. Sitopsis) had the largest values in stomatal length, girth and area, and the lowest values in stomatal width, density and index. On the contrary, the species with D genomes (Ae. Tauschii) had the highest values in stomatal width, density and index, and the lowest values in stomatal length, girth and area. The net photosynthetic rate (Pn) and stomatal conductance (Gs) were higher in species with D genomes (Ae. Tauschii) and lower in species with S genomes (Ae. Sect. Sitopsis), respectively. The net photosynthetic rate showed significant correlation with stomatal conductivity, and the changes in intercellular CO₂ content showed the opposite trend to the changes in stomatal limitation, indicating that the stomatal conductivity is the main factor limiting the photosynthetic rate. T. urartu with higher net photosynthetic ability showed higher values in stomatal conductivity and stomatal limitation, and lower values in intercellular CO₂ content. However, T. boeoticum showed the opposite trend. T. urartu had higher values of chlorophyll content in flag leaf area, while T. boeoticum had lower values, indicating that there might be higher chlorophyll content.     

普通小麦基因组最可能的4个供体的ITS1和ITS2序列及其亲缘关系

对普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）基因组（ＡＡＢＢＤＤ）最可能的供体－Ｔ．ｕｒａｔｒｔｕＴｈｕｍ．（ＡＡ）、Ｔ．ｍｏｎｏｃｃｕｍｖａｒ．ｂｏｅｏｔｉｃｕｍ（Ｂｏｉｓｓ．）ＭＫ（ＡＡ）、ＡｅｇｉｌｏｐｓｓｐｅｌｔｏｉｄｅｓＴａｕｓｃｈ．和Ａｅ．ｔａｕｓｃｈｉｉ（Ｃｏｓｓ．（ＤＤ）的核糖体ＲＮＡ基因ＩＴＳ区进行了ＰＣＲ扩增和克隆,并测定了ＩＴＳ１和ＩＴＳ２的ＤＮＡ序列,讨论和纠正了前人     

Evolutional trends of leaf stomatal and photosynthetic characteristics in wheat evolutions

According to the evolutional relationships among wheat varieties, 21 wheat varieties were chosen as research materials in this experiment to determine the evolutional trends of stomatal and photosynthetic characteristics. The results showed that as ploidy increased, the stomatal length, width, perimeter and area were found to increase. The stomatal density was found to decrease in A, B, D genomes, while no differences were found in stomatal indices among ploidies, indicating that the stomata became larger, but were still less in evolution progress. The Diploidy had the highest Pn, which was less in Tetraploidy, and the least in Hexaploidy. On the contrary, the Hexaploidy had the highest value in Fv/Fm, and the Diploidy had the highest chlorophyll content. The net photosynthetic rate had significant correlation with stomatal conductivity, while no significant relationship was found between stomatal conductivity and any stomatal characteristics, indicating that the stomatal conductivity is one of the factors limiting the photosynthetic rate, while the single stomatal characteristics is not the reason inducing the change in photosynthetic rate. The stomatal density showed significant differences among ploidy materials in A, B, D genomes, and the trend of genotype was in order of 2n > 4n > 6n; the stomatal length, width, perimeter and area showed significant differences among ploidy materials, and the trend of genotype was in order of 6n > 4n > 2n. The results indicated that the low density might induce low conductivity and low photosynthetic ability in Hexaploidy. Furthermore, the Diploidy had higher value in chlorophyll content of flag leaf area among ploidy materials in A, B and D genomes, indicating that higher chlorophyll content might be the reason for higher photosynthetic ability in Diploid wheat species.     

Studies on the origin and evolution of tetraploid wheats based on the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA

In this study, the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA in the tetraploid wheats, Triticum turgidum (AABB) and Triticum timopheevii (AAGG), their possible diploid donors, i.e., Triticum monococcum (AA), Triticum urartu (AA), and five species in Aegilops sect. Sitopsis (SS genome), and a related species Aegilops tauschii were cloned and sequenced. ITS1 and ITS2 regions of 24 clones from the above species were compared. Phylogenetic analysis demonstrated that Aegilops speltoides was distinct from other species in Aegilops sect. Sitopsis and was the most-likely donor of the B and G genomes to tetraploid wheats. Two types of ITS repeats were cloned from Triticum turgidum ssp. dicoccoides, one markedly similar to that from T. monococcum ssp. boeoticum (AA), and the other to that from Ae. speltoides (SS). The former might have resulted from a recent integression event. The results also indicated that T. turgidum and T. timopheevii might have simultaneously originated from a common ancestral tetraploid species or be derived from two hybridization events but within a very short interval time. ITS paralogues in tetraploid wheats have not been uniformly homogenized by concerted evolution, and high heterogeneity has been found among repeats within individuals of tetraploid wheats. In some tetraploid wheats, the observed heterogeneity originated from the same genome (B or G). Three kinds of ITS repeats from the G genome of an individual of T. timopheevii ssp. araraticum were more divergent than that from inter-specific taxa. This study also demonstrated that hybridization and polyploidization might accelerate the evolution rate of ITS repeats in tetraploid wheats.     

Analysis of Triticum boeoticum and Triticum urartu seed defensins: To the problem of the origin of polyploid wheat genomes

The origin of polyploid wheat genomes has been the subject of numerous studies and is the key problem in wheat phylogeny. Different diploid species have been supposed to donate genomes to tetraploid and hexaploid wheat species. To shed light on phylogenetic relationships between the presumable A genome donors and hexaploid wheat species we have applied a new approach: the comparison of defensins from diploid Triticum species, Triticum boeoticum Boiss. and Triticum urartu Thum. ex Gandil., with previously characterized Triticum kiharae defensins [T.I. Odintsova et al., Biochimie 89 (2007) 605-612]. Defensins were isolated by acidic extraction of seeds followed by three-step chromatographic separation. Isolated defensins were identified by molecular masses using MALDI-TOF mass spectrometry and N-terminal sequencing. For the first time, we have shown that T. urartu defensins are more similar to those of the hexaploid wheat than T. boeoticum defensins, although variation among samples collected in different regions of the world was revealed. Our results clearly demonstrate that T. urartu of the Asian origin contributed the A genome to polyploid wheat species.     

ITS1 and ITS2 Sequences of Four Possible Donors to Bread Wheat Genome and Their Phylogenetic Relationships

The 1TS1 and ITS2 of rDNA of four diploid species, newly Triticum urartu Thum. (AA), T. monococcum var. boeoticum (Boiss.) MK (AA),Aegilops speltoides Tausch. (BB) and Ae. taus&ii Coss. (DD), the most possible donors of A, B and D genomes to broad wheat ( T. aestivum), were amplified by PCR, cloned and sequenced. Some published sequences were discussed and rectified. The length of ITS1 sequences in four species was 221 to 223 bp, and that of 1TS2 was 216 to 217 bp. In pairwise sequence comparisons among four species, divergence ranged from 0.029 0 to 0.064 0 in ITS1, and from 0.009 3 to 0.058 0 in ITS2. Based on ITS1, ITS2 and 1TS1 + ITS2 data respectively, the same most parsimony tree that is congruent with the phylogenetic relationships was generated which was concordant with their morphological and cytological characteristics. In the trees, T. urartu and T. monococcum var. boeoticum constituted one monophyly, whereas two species of the genus Aegilops, Ac. speltoides and Ac. tauschii, fortmed another mono- phyly but with lower bootstrap value than the first clade. This study suggests that ITS region is a useful molecular marker in the studies on the origin and evolution of Triticum.     

小麦染色体组的起源与进化探讨

对小麦染色体组的起源及其进化进行了全面综述后,提出了一个新的小麦进化途径,并认为：（１）Ｔｒｉｔｉｃｕｍｍｏｎｏｃｏｃｕｍｖａｒｕｒａｒｔｕ是多倍体小麦Ａ组的原初供体,在Ａ组进入多倍体小麦后有Ｔｍｏｎｏｖａｒｂｏｅｏｔｉｃｕｍ的基因渗入;（２）Ｂ和Ｇ组的原初供体是Ｔｓｐｅｌｔｏｉｄｅｓ的Ｓ组,在该Ｓ组进入多倍体小麦后有两个进化方向,即Ｓ组结构分化形成Ｇ组和Ｓ组经外源染色体代换及重组等而进化成Ｂ组;（３）Ｔｔｕｒｇｉｄｕｍ和Ｔｔｉｍｏｐｈｅｖｉ都是来自Ｔｓｐｅｌｔｏｉｄｅｓ为母本与Ｔｍｏｎｏｖａｒｕｒａｒｔｕ杂交后并双二倍化而形成的原初四倍体小麦（ＳＳＡＡ）,并由它分别经遗传渗入和结构分化而成;（４）Ｔｚｈｕｋｏｖｓｋｙｉ是Ｔｔｉｍｏｐｈｅｖｉ作母本与Ｔｍｏｎｏｖａｒｂｏｅｏｔｉｃｕｍ杂交并双二倍化而形成,故它具有分别来自Ｔｍｏｎｏｖａｒｕｒａｒｔｕ和Ｔｍｏｎｏｖａｒｂｏｅｏｔｉｃｕｍ的两类Ａ组;（５）Ｔａｅｓｔｉｖｕｍ的Ｄ组来自Ｔｔａｕｓｃｈｉ;（６）无论Ａ组、Ｂ组、Ｄ组、Ｇ组在进入多倍体小麦后均有相当分化,同时在其供体种中也有一定分化。     

Electrophoretic survey of seedling esterases in wheats in relation to their phylogeny

Summary Evolutionary and ontogenetic variation of six seedling esterases of independent genetic control is studied in polyploid wheats and their diploid relatives by means of polyacrylamide gel electrophoresis. Four of them are shown to be controlled by homoeoallelic genes in chromosomes of third, sixth and seventh homoeologous groups.The isoesterase electrophoretic data are considered supporting a monophyletic origin of both the primitive tetraploid and the primitive hexaploid wheat from which contemporary taxa of polyploid wheats have emerged polyphyletically and polytopically through recurrent introgressive hybridization and accumulation of mutations. Ancestral diploids belonging or closely related to Triticum boeoticum, T. urartu, Aegilops speltoides and Ae. tauschii ssp. strangulata are genetically the most suitable genome donors of polyploid wheats. Diploids of the Emarginata subsection of the section Sitopsis, Aegilops longissima s.str., Ae. sharonensis, Ae. searsii and Ae. bicornis, are unsuitable for the role of the wheat B genome donors, being all fixed for the esterase B and D electromorphs different from those of tetraploid wheats.     

Biochemical data bearing on the relationship between the genome of Triticum urartu and the A and B genomes of the polyploid wheats

To determine whether the Triticum urartu genome is more closely related to the A or B genome of the polyploid wheats, the amino acid sequence of its purothionin was compared to the amino acid sequences of the purothionins in Triticum monococcum, Triticum turgidum, and Triticum aestivum. The residue sequence of the purothionin from T. urartu differs by five and six amino acid substitutions respectively from the alpha 1 and alpha 2 forms coded for by genes in the B and D genomes, and is identical to the beta form specified by a gene in the A genome. Therefore, the T. urartu purothionin is either coded by a gene in the A genome or a chromosome set highly homologous to it. The results demonstrate that at least a portion of the T. urartu and T. monococcum genomes is homologous and probably identical. A variety of other studies have also shown that T. urartu is very closely related to T. monococcum and, in all likelihood, also possesses the A genome. Therefore, it could be argued that either T. urartu and T. monococcum are the same species or that T. urartu rather than T. monococcum is the source of the A genome in T. turgidum and T. aestivum. Except for Johnson's results, our data and that of others suggest a revised origin of polyploid wheats. Specifically, the list of six putative B genome donor species is reduced to five, all members of the Sitopsis section of the genus Aegilops.