拟南芥低叶绿素荧光LCF3基因的克隆与功能分析

叶绿素荧光动力学技术在测定叶片光合作用中光系统对光能的吸收、传递、耗散、分配等方面具有独特的作用。本研究借助叶绿素荧光成像仪, 从拟南芥EMS诱变突变体库中筛选到一株低叶绿素荧光突变体lcf3-1 (lower chlorophyll fluorescence 3-1)。遗传分析表明lcf3-1突变体为单基因隐性突变。突变基因图位克隆结果显示LCF3是PsbW的等位基因。另外, LCF3基因的T-DNA插入突变体及功能回补转基因植物的叶绿素荧光分析结果, 均证明LCF3基因突变导致拟南芥叶绿素荧光F_v/F_m值降低。进一步实验证明LCF3蛋白定位于叶绿体, 且LCF3基因在植株中普遍表达。PsbW蛋白可能细微调整PSII-LHCII超复合体的组装及稳定。     

Impact of wheat-Leymus racemosus added chromosomes on wheat adaptation and tolerance to heat stress

Adaptation of wheat (Triticum aestivum L.) to high temperatures could be improved by introducing alien genes from wild relatives. We evaluated the responses of wheat-Leymus racemosus chromosome introgression lines to high temperature to determine their potentiality for developing improved wheat cultivars. Introgression lines and their parent Chinese Spring were evaluated in a growth chamber at the seedling stage and in the field at the reproductive stage in two heat-stressed environments in Sudan. Optimum and late planting were used to ensure exposure of the plants to heat stress at the reproductive stage. The results revealed the impact of several Leymus chromosomes in improving wheat adaptation and tolerance to heat. Three lines possessed enhanced adaptation, whereas two showed high heat tolerance. Two addition lines showed a large number of kernels per spike, while one possessed high yield potential. Grain yield was correlated negatively with the heat susceptibility index, days to heading and maturity and positively with kernel number per spike and triphenyl tetrazolium chloride assay under late planting. The findings suggest that these genetic stocks could be used as a bridge to introduce the valuable Leymus traits into a superior wheat genetic background, thus helping maximize wheat yield in heat-stressed environments.     

Plant regeneration from cotyledons and embryonic axes in apple: Sites of reaction and effect of pre-culture in the light

Summary Mature embryos of the cultivar Gloster were isolated from seeds of fruits stored at 1 °C. Embryonic axes and wounded cotyledons were pre-cultured in the light for 0 to 6 days on embryo growth medium (experiment 1) or for 0 to 24 hours on shoot induction medium (experiment 2). After a subsequent dark period of 3 weeks for induction of organogenesis, explants were cultured in the light on shoot elongation medium. Up to 95% of the embryos formed shoots, while the mean shoot number per embryo reached 6.4. Regeneration frequency was influenced by the type of explant: embryonic axes reacted better (75%) than the tops of the cotyledons (29%) or wounds on the cotyledons (9%). The mean total shoot number per embryo comprised means of 2.3 for embryonic axes, 1.8 for tops of cotyledons and 2.3 for wounds on cotyledons (experiment 1). Wounds on the proximal part of the cotyledon reacted much better than those on other parts. Shoot differentiation was highest when cotyledons were cultured with the adaxial side on the medium, suggesting that there exist gradations within the cotyledon for regeneration capacity (experiment 2). The number of shoots per reactive site was not normally distributed, while a high frequency of 6 or more shoots per reactive site was found (experiment 1). A pre-culture of more than 24 hours in the light seemed to have negative effect, but there was no clear evidence whether this was a light and/or a medium effect.     

Identification of QTLs for fruit quality traits in Japanese apples: QTLs for early ripening are tightly related to preharvest fruit drop

Many important apple (Malus × domestica Borkh.) fruit quality traits are regulated by multiple genes, and more information about quantitative trait loci (QTLs) for these traits is required for marker-assisted selection. In this study, we constructed genetic linkage maps of the Japanese apple cultivars ‘Orin’ and ‘Akane’ using F₁ seedlings derived from a cross between these cultivars. The ‘Orin’ map consisted of 251 loci covering 17 linkage groups (LGs; total length 1095.3 cM), and the ‘Akane’ map consisted of 291 loci covering 18 LGs (total length 1098.2 cM). We performed QTL analysis for 16 important traits, and found that four QTLs related to harvest time explained about 70% of genetic variation, and these will be useful for marker-assisted selection. The QTL for early harvest time in LG15 was located very close to the QTL for preharvest fruit drop. The QTL for skin color depth was located around the position of MYB1 in LG9, which suggested that alleles harbored by ‘Akane’ are regulating red color depth with different degrees of effect. We also analyzed soluble solids and sugar component contents, and found that a QTL for soluble solids content in LG16 could be explained by the amount of sorbitol and fructose.     

野生二粒小麦导入普通小麦的抗白粉病基因MlWE29分子标记定位

小麦白粉病是严重影响小麦生产的重要病害之一,培育和应用抗病品种是有效控制和减少病害的最经济有效的方法。野生二粒小麦是硬粒小麦和普通小麦的四倍体野生祖先种,是小麦抗病性遗传改良的重要基因资源。本研究利用来自以色列的野生二粒小麦WE29与普通小麦杂交,再用普通小麦连续回交和自交,育成高抗白粉病(Blumeria graminis f. sp. tritici)小麦新品系3D258(系谱为燕大1817/WE29//5*87-1, BC₄F₆)。将3D258和高感小麦白粉病的普通小麦品种薛早配制杂交组合,对其F₁、F₂代分离群体和F₃代家系进行白粉病抗性鉴定和遗传分析。结果表明3D258携带抗白粉病显性单基因,暂命名为MlWE29。利用集群分离分析法(BSA)和分子标记分析,发现6个SSR标记(Xgwm335、Xgwm213、Xgwm639、Xwmc415、Xwmc289和Xwmc75)和5个EST-STS标记(BE494426、BE442763、CD452476、BE445282和BE407068)与抗白粉病基因MlWE29连锁。利用中国春缺体-四体系、双端体系和缺失系将抗白粉病基因MlWE29标记物理定位于5BL染色体的0.59–0.79区域。这一普通小麦抗白粉病种质资源的创制及其连锁分子标记的建立为小麦抗病基因分子标记辅助选择、基因积聚和分子育种提供了新的物质基础。     

羽衣甘蓝自交不亲和与自交亲和系种子萌发期DNA甲基化的动态变化

自交不亲和系植株的种子往往出现退化现象,为了研究种子的退化现象是否与甲基化相关,因此本文采用甲基化敏感扩增多态性(methylation sensitive amplification polymorphism, MSAP)技术,以羽衣甘蓝自交不亲和系9^#种子、自交亲和系14^#种子为研究对象,对其生长发育过程种子基因组DNA甲基化水平变化情况进行研究。采用改良的CTAB法提取种子萌发不同时期DNA,然后通过MSAP分析、统计扩增条带,比较二者之间的差异。对9^#种子DNA甲基化状态分析表明,萌发前期(0~2 d)发生甲基化位点数目持续增多,但萌发后期(2~8 d)发生去甲基化的数目大量增加,整个萌发期去甲基化位点数目是甲基化位点数目的11倍,说明9^#种子萌发过程中DNA甲基化修饰是基因表达的重要调控方式之一;在相同发育时期,9^#在总甲基化、全甲基化、半甲基化水平上均不同程度高于14^#。随着种子的萌发,9^#全甲基化水平明显上升,半甲基化水平几乎不变,而14^#变化趋势与9^#相反,半甲基化水平明显上升,全甲基化水平几乎不变.     

Restrictions of the efficiency of haploid plant production in apple cultivar Idared,through parthenogenesisin situ

Summary For haploid plant production in Idared apple, parthenogenesisin situ was induced by irradiated pollen. The most important restrictions were seed set, embryo germination and green plant recovery from germinated embryos. The efficiency of green plant production was strongly influenced by irradiation dose, picking time and quality of the irradiated pollen. 250 Gy pollen was more efficient for green plant production than 500 Gy pollen. Using 250 Gy pollen with reduced germination capacity resulted in a five-fold lower green plant yield. Delaying picking of the fruits (140 instead of 70 days after pollination) had a significant positive effect. Up to 20 plants per 1000 pollinated flowers, showing no expression of a marker gene present in the pollen parent, were obtained in some treatments. Induction efficiency was high when seed weight was high. This offers the possibility of selecting seeds with the best chances for haploid plant production. Heavy seeds generally contained visual normal developed embryos, which germinated better than those from lighter seeds and gave normal plant development. Delayed picking increased the proportion of large seeds, suggesting that pollination with irradiated pollen retards embryo development. From treatments with irradiated pollen of poor quality more small seeds were harvested.     

转TiERF1-RC7双价基因小麦的鉴定及其全蚀病抗性

为探讨TiERF1和RC7基因对小麦抗全蚀病的防御反应,本研究对转TiERF1-RC7双价基因小麦进行了分子检测以及全蚀病抗性的室内和田间鉴定。结果表明,转入的TiERF1和RC7基因在转基因小麦中可以遗传和转录;与受体扬麦18相比,5个转TiERF1-RC7小麦株系在整个生育期抗病性显著提高,苗期的全蚀病严重度在10%以下,成熟期的白穗率在13%以下,而扬麦18的严重度为62.98%,白穗率为26.09%。电子显微镜观察结果表明抗病转基因小麦根表的全蚀病原菌菌丝数量及生长势明显低于感病材料。上述结果说明,转入的TiERF1和RC7基因抑制了全蚀病原菌的侵染及在转基因小麦中繁殖,进而提高了转基因小麦对全蚀病的抗性。     

异源表达棉花GhPRP5基因增强了拟南芥对盐和ABA的敏感性

富含脯氨酸的蛋白(proline-rich proteins, PRPs)代表一类富含脯氨酸和羟脯氨酸的细胞壁结构蛋白质,最先在伤害诱导的胡萝卜贮藏根中被发现。越来越多的证据显示这类蛋白在应答多种生物和非生物胁迫中起作用。我们之前从棉花cDNA文库中分离了一个命名为GhPRP5的编码富含脯氨酸蛋白的基因,为研究其功能,构建了GhPRP5的过量表达载体,转化拟南芥,获得GhPRP5高表达的8个株系的纯合体。在正常培养条件下,转基因株系和野生型种子的萌发率一致,但盐胁迫和ABA处理显著抑制了转基因拟南芥株系种子的萌发。盐胁迫条件下野生型的绿苗率明显高于转基因拟南芥株系,与正常生长条件相比,ABA处理抑制转基因拟南芥主根伸长的程度更大。利用Quantitative RT-PCR技术分析几个胁迫相关标记基因的表达情况表明,盐和ABA诱导了RD29A、RD29B和KIN1的表达,但诱导水平在转基因株系和野生型中不一样,说明GhPRP5参与调控拟南芥胁迫相关标记基因的表达,但具体参与的胁迫应答信号传导途径仍需进一步研究。     

利用斑茅cDNA芯片研究甘蔗受黑穗病菌侵染后基因差异表达

为了解甘蔗抗黑穗病的分子机制,利用斑茅干旱胁迫cDNA芯片检测了甘蔗受黑穗病菌胁迫后的基因差异表达。经杂交,在cDNA芯片的3 860个模板中,有效差异表达(Ratio值≥2.0或≤0.5)的基因为101个,其中上调55个,下调46个。部分基因的定量PCR验证表明,芯片杂交结果可靠。上调表达基因经测序、冗余序列剔除,一共获得36个unique ESTs。已知功能的22个上调表达基因,涉及多条生理代谢途径,如光合作用、离子转运和核酸代谢途径;以及多种分子水平的进程,如基因转录、蛋白质合成与修饰以及细胞信号转导等。另外,还检测到14个未知功能基因。结果表明,甘蔗对黑穗病的抗性具有复杂的机制。本研究为解释甘蔗受黑穗病菌胁迫后的基因差异表达及其网络构建奠定了基础,还可以为今后系统研究甘蔗对生物与非生物胁迫的响应机制提供技术支持。     

Recent advances on the use of natural and safe alternatives to conventional methods to control postharvest gray mold of table grapes

Gray mold, caused by Botrytis cinerea, is the main postharvest decay of table grapes. It can develop in the vineyard and spread rapidly among berries after harvest, during long distant transport, cold storage and shelf-life. In conventional agriculture, bunches are sprayed with fungicides after flowering, at pre-bunch closure, at veraison, and later, depending on the time of harvest. Harvested bunches are usually stored in the presence of sulfur dioxide. However, the use of synthetic fungicides and of sulfur dioxide is not allowed on organic grapes and the study of alternative methods to control postharvest decay has developed over several decades, along with the demand for safer storage methods. This review summarizes the results published in the field within the last 5 years (2006-2010). We can group these approaches as follows: (i) biocontrol agents; (ii) natural antimicrobials; (iii) GRAS type decontaminating agents; and (iv) physical means. Two biocontrol agents, Muscodor albus and Hanseniaspora uvarum, have shown equal or better effectiveness than conventional methods to control gray mold of table grapes in laboratory scale experiments. Currently, the bottleneck for the commercial use of biocontrol agents is that the registration process is comparable to that of fungicides, with similar costs but often with a narrower market. This delays their transition from experimental to practical use. Natural antimicrobials, such as salts, chitosan, and plant extracts, have demonstrated good results and often have been applied in various scales. Several GRAS-classified sanitizers have been tested to extend postharvest storage of table grapes, including acetic acid, electrolyzed oxidizing water, ozone, and ethanol. Physical technologies involving variations in temperature, UV-C irradiation, pressure or changing atmospheric composition, are all postharvest practices which require significant adaptation by an industry which is accustomed to minimal intervention during harvest. Overall, the use of ozone and of calcium chloride are two promising examples of treatments that are beginning to be adopted on a commercial scale. The requirements for the optimal treatment of grapes against gray mold before harvest or during storage are summarized.     

The combined effect of plant stature and maturity on the response of wheat and triticale accessions to Septoria tritici

Summary The relationships between percent pycnidia coverage on the four uppermost leaves (PCD), plant height (PHT) and days to heading (HED) were evaluated for 21,000 wheat and triticale accessions tested in artificially inoculated (with fixed combination of S. tritici isolates) field nurseries over 8 trial years. A general Linear Model procedure (GLM) estimated Septoria severity using two correlative models: model 133-1 Year and model II–PCD=b₁PHT+b₂HED+C. The regression coefficients for PHT and HED in the two models were –0.54 and –0.40, respectively, with a R²=0.80^** and R²=0.29^** for model I and model II, respectively. The predicted cultivar best fitted to the model would be characterized as a semidwarf (PHT=115 cm) with an early-moderate maturity (HED=95 days to heading). The estimated mean percent pycnidial coverage for the two models over the 8 trial years was 40.8%. The performance of a group of 38 cultivars replicated yearly during the 8 trial years was assessed relative to model I. The deviation of each cultivar from the model was calculated using two functions: a) Sum Relative Serial Deviation (SRSD) and b) Total Relative deviation (TRD), in addition to Standard errors (SE). The proposed analytical protocol enabled identification of cultivars which expressed consistent yearly deviation (from the model) in host response combined with low-moderate mean pycnidial coverage (±30%). Such cultivars may possess a more stable type of genetic protection against the adverse effects of septoria tritici blotch.     

Genetic structure and diversity of the wild Ussurian pear in East Asia

The Ussurian pear is the most important cultivated pear in the northern part of China. Cultivated Ussurian pears are considered to have derived from Pyrus ussuriensis Maxim. which is native to the northeast of China. In Japan, two varieties of P. ussuriensis, P. ussuriensis var. aromatica and var. hondoensis are native to the northern area and the central area of the main island respectively. In order to reveal the origin of Pyrus ussuriensis var. aromatica distributed in the northern area of main island of Japan, more than 40 explorations have been performed in Japan and in China, and more than 30 natural habitats were recognized. These natural habitats are at risk of extinction because of human development and forest degradation caused by climate change. Population structure and genetic diversity of P. ussuriensis in China and P. ussuriensis var. aromatica in Japan have been investigated using both morphological and molecular markers in order to define appropriate conservation units, and to provide a good focus for conservation management. Distant evolutionary relationships between P. ussuriensis Maxim. in China and P. ussuriensis var. aromatica in Japan inferred from population genetic structure and phylogenetic analysis are also discussed.     

基于EST-PCR的簇毛麦染色体特异分子标记筛选及应用

为定位、转移和利用簇毛麦有益基因, 通过花粉辐射, 获得一批包括小麦-簇毛麦易位染色体的异染色体系。为了鉴定这批材料中的簇毛麦染色体身份, 根据水稻、小麦的EST序列合成了240对STS引物, 其中34对引物在普通小麦中国春与簇毛麦间存在多态性;进一步对亲本及簇毛麦二体异附加系进行PCR扩增分析, 标记CINAU32_-300可追踪簇毛麦1V染色体, 标记CINAU33_-280、CINAU34_-510、CINAU35_-1100、CINAU36_-380和CINAU37_-400可追踪簇毛麦2V染色体, 标记CINAU38_-250可追踪簇毛麦3V染色体, 标记CINAU39_-950和CINAU40_-800可追踪簇毛麦4V染色体, 标记CINAU41_-745和CINAU42_-1050可追踪簇毛麦5V染色体, 标记CINAU44_-765和CINAU45_-495可追踪簇毛麦7V染色体。加上本室已开发的2个6V染色体特异标记, 用这些簇毛麦特异分子标记鉴定辐射诱导材料的部分回交后代, 选育出小麦背景中只包含单条簇毛麦染色体的整套1V至7V染色体系, 同时有18条易位染色体的簇毛麦身份得到确定, 表明这些标记可以用来快速检测普通小麦背景中的簇毛麦染色体或染色体片段。     

Leaf thickness and response of leaf photosynthesis to water stress in soybean varieties

Summary A comparative study of photosynthetic response to soil moisture was conducted with soybean varieties (Harosoy, Norin No. 1 and Hogyoku). When the plants were grown in May before the rainy season under a high radiation level, per unit leaf area the photosynthetic rate of Harosoy leaves was significantly higher throughout the entire range of soil water potentials and leaf water potentials than that of Norin No. 1 leaves. The high photosynthetic rate of Harosoy under non-stress conditionsaand mild water stress was associated with both the high specific leaf weight and the high leaf areal nitrogen content. The photosynthetic response of the Harosoy leaves developed during rainy season under a low radiation level, however, was similar to that of Norin No. 1; no significant difference was observed between these two varieties in the specific leaf weight and in the leaf areal nitrogen content.On the other hand, Hogyoku cultivated especially along the footpaths between the paddy field was very sensitive to water stress, closing the stomata at higher soil water potentials. Under non-stress conditions, as compared with per unit leaf nitrogen content, however, Hogyoku performed higher leaf photosynthesis even with a thinner leaf than Harosoy.     

高温胁迫对水稻灌浆结实期叶绿素荧光、抗活性氧活力和稻米品质的影响

选用西农优30和富优1号两个水稻品种, 在灌浆结实期利用人工气候室进行高温处理, 测定叶绿素含量、叶绿素荧光参数、抗活性氧活力的动态及成熟稻米的营养品质。结果表明, 高温胁迫下水稻叶片叶绿素含量降低, 叶绿素荧光参数初始荧光(F_o)明显上升, PS II的潜在活性(F_v/F_o)和PS II的最大光能转化效率(F_v/F_m)均低于对照, 呈下降趋势; 灌浆结实后期F_v/F_o和F_v/F_m的下降速率最快; 灌浆结实后期抗氧化活力低于对照。同时, 高温胁迫下的成熟稻米总淀粉及直链淀粉含量降低, 蛋白质含量升高, 垩白度上升而千粒重下降。高温胁迫下光合作用的原初反应被抑制, 抗活性氧活力降低, 水稻叶片功能期缩短, 可能是稻米品质降低的重要原因。     

小麦TaFT基因编码区序列多态性及其与开花期的关系

春化基因VRN-B3是小麦开花素基因TaFT,为探索该基因在品种间的保守性及其与小麦开花早晚的关系,根据TaFT基因序列(GenBank accession No.: DQ890162)设计特异PCR引物,扩增了13个品种中该基因的编码区。通过测序和序列比对,发现不同品种间该基因编码区的DNA序列存在多态性,序列翻译发现5个品种的表达产物FT蛋白发生变异。利用中国春的非整倍体材料将TaFT基因定位在7BS染色体上。参考品种的冬春性及开花时间,推测冬性品种正常的FT蛋白(同DQ890162翻译的氨基酸序列一致)可加速开花,FT蛋白变异则延迟开花;春性品种的FT蛋白变异与否对开花期影响不大,推测TaFT基因的效应可能被春性品种的显性春化基因所掩盖。     

The effect of temperature and other factors on chlorophyll a fluorescence and the lower oxygen limit in apples (Malus domestica)

The effects of temperature, scan interval and rate of oxygen (O₂) decline on pulse frequency modulation (PFM)-based minimum fluorescence (F_α) and the F_α-based lower oxygen limit (LOL) were investigated using ‘Honeycrisp’ apples (Malus × domestica Borkh). The effects of temperature and hypoxic stress on pulse amplitude modulation (PAM) fluorescence parameters were also investigated. The PFM scan interval had no effect on the F_α baseline, but increases in the scan interval decreased the low-O₂-induced fluorescence spike intensity (ΔF_α). Temperature negatively correlated with the F_α baseline while ΔF_α °C⁻¹ was greater at lower than at higher temperatures. When using a PAM fluorometer, the minimum fluorescence (F_o), and to a lesser extent the maximum fluorescence (F_m), were similarly affected by temperature as was F_α. Temperature altered the LOL in fruit. Apples stored at 20, 10, 3.5 and 0 °C spiked at 0.72, 0.33, 0.22 and 0.08 kPa O₂, respectively, under a rapid O₂ decline (i.e., 20.9 to <1 kPa O₂ in ≈5–6 h). Although the low-O₂ F_α spike apex values did not change with temperature, the spike intensity increased with temperature due to a reduced fluorescence baseline. A slower O₂ decline rate produced slightly higher LOL and lower spike intensity values. In conclusion, temperature and rate of O₂ decline affected the low-O₂-induced PFM fluorescence spike intensity as well as the LOL, while the PFM scan interval affected the spike intensity.     

Genetic analysis of drought stress response in rapeseed (Brassica campestris and B. napus). I. Assessment of environments for maximum selection response in grain yield

Summary Expectations of yield improvement in environments where drought was the major environmental factor limiting yields were studied in two species of rapeseed. Selection for yield in a drought stressed environment was predicted to be a more efficient selection stategy for yield improvement in dryland situations than selection in a more optimal environment, or selection based on a drought response index. The results indicate that selection for yield in a stressed environment are expected to lead to genetic advances in yield under optimal conditions as well as in a drought index. Selection under well watered conditions, on the other hand, was also expected to lead to correlated increases in yield in droughted environments but to decreases in the drought index. These results were found in both species of rapeseed grown in different water stress situations.The genetic advance in a drought response index was predicted to be greater in B. napus and marginally less in B. campestris if selection was practised for yield in a stressed environment rather than direct selection for the drought index. This was due to the higher heritability estimates in the stressed environments and the positive genetic correlations with yield.     

Geneticanalysis of drought stress response in rapeseed (Brassica campestris and B. Napus). II. Yield improvement and the application of selection indices

Summary The expected improvement in grain yield if selection was made for plant characters measured between flowering and maturity was evaluated in populations of rapeseed (Brassica campestris and B. napus) grown in a droughted environment. Drought was commenced at flowering in each species and measurements were made on plants at the commencement of the drought stress, during the stress treatment and at crop maturity.Substantial genetic and phenotypic variation was observed in yield as well as the different morphophysiological determinants of yield. In B. campestris no single parameter was found to be a suitable alternative selection criterion to yield since the correlated responses in yield if selection was for another character was lower than if selection was for yield alone. By the use of a selection index however, joint selection for yield, as well as harvest index, 1000 seed weight and seeds per pod, was expected to be 20% more effective than direct selection for yield under drought. In the B. napus population direct selection for flowering time or for harvest index was predicted to result in a genetic advance in yield equal to or greater than that obtained by direct selection for yield, whereas joint selection for flowering time and yield should result in a 16% greater yield increase. Selection for increased yield in these populations grown in droughted conditions is discussed.