甘蔗热带种金属硫蛋白家族基因的克隆及响应重金属胁迫的表达分析

金属硫蛋白是一类富含巯基的低分子量蛋白,在植物的重金属解毒及细胞氧化还原调控等方面起重要的作用。本研究以甘蔗热带种Badila组培苗为材料,分别测定了其在CdCl2、ZnSO4和CuCl2水溶液培养条件下地上部和地下部的重金属含量,结果显示其对上述3种重金属有较强的耐受与富集能力。继而克隆了ScMT1(登录号为KJ504373)、ScMT2-1-5 (登录号为MH191346)和ScMT3 (登录号为KJ5043704) 3个金属硫蛋白家族基因,它们分别属于植物MT亚家族中的MT1、MT2和MT3型基因。ScMT1含有1个内含子和2个外显子,开放阅读框(Open Reading Frame, ORF)长228 bp,编码75个氨基酸; ScMT2-1-5含有2个内含子和3个外显子, ORF长246 bp,编码81个氨基酸; ScMT3含有1个内含子和2个外显子, ORF长198 bp,编码65个氨基酸。RT-qPCR显示, Cd2+胁迫下,在甘蔗地上部和地下部, ScMT2-1-5均连续显著上调表达,而ScMT1的上调应答出现延迟。ScMT3在地上部的上调应答出现延迟,在地下部呈"扬–...     

用AFLP分子标记技术鉴别甘蔗(Saccharum officinarum L.)品种

本研究采用AFLP技术,从64对引物中筛选出5对带型分布均匀、多态性丰富且分辨能力强的引物,对两个在生产上广泛推广应用的甘蔗品种的AFLP指纹图谱进行分析,计算了两品种间的遗传相似性和遗传距离.结果表明,5对引物在2个甘蔗品种间均存在显著的差异,其中多态性位点占总扩增位点的10.2%,区分率达100%.研究结果对生产上应用AFLP技术鉴定甘蔗品种的真伪性提供了依据.本文对甘蔗AFLP分析体系的关键技术以及银染技术的安全性和可靠性进行了讨论.     

甘蔗（Saccharum officinarum L.）品种遗传差异的AFLP分子标记分析

采用AFLP技术,从64对引物中筛选出5对带型分布均匀、多态性丰富且分辨能力强的引物,对两个在生产上广泛推广应用的甘蔗品种的AFLP指纹图谱进行分析,计算了两品种间的遗传相似性和遗传距离。结果表明,5对引物在2个甘蔗品种间均存在显著的差异,其中多态性位点占总扩增位点的10.2%,区分率达100%。这对应用AFLP技术鉴定甘蔗     

Irrigation Regimes on Utilization of Nutrients, Yield and Quality of Sugarcane (Saccharum officinarum L.)

With a view to study the influence of irrigation regimes at critical stages of sugarcane an investigation was carried out at Agricultural Research Station, Tamil Nadu Agricultural University, Bhavanisagar, India during Mid-late season (March–April) using Co 6304 as test variety. Treatments consisted of eight irrigation regimes created by imposing two levels of irrigation 0.75 and 0.5 IW/CPE ratio at critical stages (tillering, grand growth and maturity). The results revealed that irrigating sugarcane at 0.75 IW/CPE ratio at tillering and growth and 0.5 IW/CPE at maturity phase recorded highest nutrient uptake, cane yield (139 t/ha) with 10.6 % CCS consuming 1791 mm of water the highest water use efficiency of 77.3 kg/ha mm.     

Potassium and Silicon Improve Yield and Juice Quality in Sugarcane (Saccharum officinarum L.) under Salt Stress

Soil salinity is a major abiotic stress which adversely affects the yield and juice quality in sugarcane. However, the mineral nutrient status of plant plays a crucial role in increasing plant tolerance to salinity. We investigated the effects of K and/or Si on plant growth, yield and juice quality in two sugarcane genotypes differing in salinity tolerance. Addition of K and Si significantly (P ≤ 0.05) increased K and Si concentrations and decreased the accumulation of Na⁺ in plants under salt stress. Cane yield and yield attributes were significantly (P ≤ 0.05) higher where K and Si were added. Juice quality characteristics like Brix (% soluble solids in juice), Pol (% sucrose in juice), commercial cane sugar (CCS) and sugar recovery in both sugarcane genotypes were also significantly (P ≤ 0.05) improved with the supplementation of K and Si. For most of the growth parameters, it was found that K either alone or in combination with Si was more effective to alleviate salt stress in both sugarcane genotypes than Si alone. Moreover, the beneficial effects of K and Si were more pronounced in salt sensitive genotype than in salt tolerant genotype. The results suggested that K and Si counteracted the deleterious effects of high salinity/sodicity in sugarcane by lowering the accumulation of Na⁺ and increase in K⁺ concentration with a resultant improvement in K⁺/Na⁺ ratio which is a good indicator to assess plant tolerance to salinity.     

干旱条件下单用草甘膦或添加营养元素作为甘蔗化学催熟剂的效应

中熟甘蔗品种桂糖15号在生长后期分别用草甘膦、 草甘膦+KH2PO4、 草甘膦+H3BO 3和草甘膦+KH2PO4+H3BO3进行叶面喷施。 结果表明： 在严重干旱的条件下, 单 用草甘膦处理对甘蔗生长和蔗茎糖分积累都不利, 而添加营养元素的处理, 尤其是草甘膦 +H3BO3处理, 可缓解草甘膦对生长代谢的抑制效应, 使叶片Mg2+-ATP酶和NADP -     

Performance of Sugarcane (Saccharum officinarum L.) Under Surface Drip, Sub Surface Drip (Biwall) and Furrow Methods of Irrigation

Field experiments were conducted at the Sugarcane Breeding Institute, Coimbatore, India during 1989–93 to study the productivity and water use efficiency of sugarcane under different methods of irrigation. The results showed that there could be a 44 % saving in water with higher water use efficiency in drip irrigation compared to conventional furrow irrigation. Among drip irrigation systems in the plant crop, sub surface drip (Biwall) at 40/140 cm spacing recorded a significantly higher number of millable canes, cane length and single cane weight compared to either conventional furrow irrigation or furrow irrigation based on IW/CPE (Irrigation Water Cumulative Pan Evaporation) ratio. In ratoon crop, Biwall irrigation at 60/120 cm gave significantly higher cane length and single cane weight compared to any other methods of irrigation. However, cane and sugar yields in Biwall irrigations were on par with conventional furrow irrigation but superior to either surface drip or furrow irrigation based in IW/CPE ratio.     

Cytological studies of 2n male gamete formation in sugarcane, Saccharum L.

The germplasm for modern sugarcane cultivars (Saccharum spp. hybrids)has been derived principally from S. officinarum (2n = 80), and S. spontaneum (2n = 40 to 128). Diploid gamete formation has been significant in developing cultivated sugarcane, but the cytological basis for the processes involved is not clearly understood. This research investigated microsporogenesis in nine clones of Saccharum spp. Hybrids and in S. officinarum and S. spontaneum. Diploid gamete formation occurred in all 11 lines, but was least frequent in S. spontaneum and S. officinarum which produced 0.5% and 0.8%2n gametes, respectively. In the hybrid lines, 2n gametes were formed infrequencies ranging from 0.9% to 4.4%. Cytological evidence was obtained for dyad and triad formation during microsporogenesis. Detailed analysis of chromosome behaviour at meiosis indicated that 2n male gamete formation is probably attributable to the absence of cytokinesis rather than a combination of asynchrony and non-disjunction. The clones were ranked on the basis of the frequencies with which they formed 4 × 1n microspores and the data were analysed using χ ² tests for homogeneity. These established that theSaccharum spp. hybrids could be designated as either ‘high’ or ‘low’ frequency haploid gamete producers. Conversely, the latter group, which formed diploid gametes most frequently (2.2%–4.4%), can be described as high frequency diploid gamete producers. The identification of clones most frequently forming diploid gametes may facilitate the more rapid recovery of desirable sugarcane genotypes because such clones could be selected for preferential use in clonal improvement. This revised version was published online in July 2006 with corrections to the Cover Date.     

Studies on Iron Chlorosis of Sugar Cane (Saccharum officinarum L.) at Metahara, Ethiopia: Soil and Plant Characterisation and Efficiency of Different Iron Sources

In order to understand the nature and causes of chlorosis at Metahara Sugar Estate (8°53′N and 39°52′E) in Ethiopia, soil profile samples were collected from a ratoon field that exhibited chlorosis. From the same site, the first ratoon sugar cane leaf blades (+ 3) were sampled on the basis of the degree of chlorosis, and thus categorised into green, medium green, medium yellow and yellow. Soil sample analysis indicated that DTPA-extractable Fe, Mn and Cu were adequate for normal plant growth in spite of an increase in CaCO₃ content. Moreover, the chlorotic leaves contained higher iron content than the green ones. Consequently, it is concluded that iron chlorosis of sugar cane at Metahara could be due to physiological inactivation of iron in soils and the plant. A field experiment was also conducted using sugar cane variety CO-449 in order to select the most effective iron source to control iron chlorosis of the plant. The plots received the treatments of FeSO₄, EDDHA and DTPA at three different levels each as foliar sprays. The untreated plots were used as control. The chemicals were applied on the first ratoon 3 weeks after harvest, and repeated a week after the first spray. Twenty millable canes from the middle two rows were sampled at 12 months for yield and juice quality determinations. Iron sulphate spray at 15 kg ha^-1 increased cane length and commercial cane sugar by 33 and 44 % over the control, respectively. The highest cane yield was secured by spraying DTPA at 1.80 kg ha^-1 even though its recoverable sugar percentage was low. Not all sources of iron improved juice quality. Since FeSO₄ at 15 kg ha^-1 was equally effective as the 30 kg ha^-1 FeSO₄, applications, the need to consider other than the currently used rate in the Estate is timely and is very important.     

甘蔗和斑茅Ⅲ型过氧化物酶基因（SoPOD-1和SaPOD-1）cDNA克隆与分析

研究旨在通过克隆甘蔗和斑茅Ⅲ型过氧化物酶基因(Prxs),分析比较2个物种Prxs基因差异,探讨2个物种Prxs功能差异的遗传基础。采用同源克隆方法,以高粱Prxs基因序列为探针,克隆获得甘蔗和斑茅Prxs基因。本研究从斑茅和甘蔗中成功克隆获得了4个Prxs基因cDNA序列,其中斑茅2个基因长度均为1083 bp,存在一个差异位点,甘蔗2个基因长度均为1084 bp,存在11个碱基差异。4个基因开放阅读框均为996 bp,编码331个氨基酸。蛋白质保守结构域分析表明,克隆获得的4个基因均具有Prxs典型保守结构域,为目标基因。系统进化分析表明,克隆获得的基因与高粱、玉米和水稻的同源基因相似性最高,在一些双子叶植物,甚至裸子植物也能找到其同源基因,且氨基酸序列的相似程度与物种分化的程度基本一致,可供植物分类作参考。该试验从斑茅和甘蔗中成功克隆获得了4个Prxs基因cDNA序列,为深入研究Prxs基因在不同植物中功能差异提供了一定的参考。     

Identification and characterization of intergeneric hybrid of commercial sugarcane (Saccharum spp. hybrid) and Erianthus arundinaceus (Retz.) Jeswiet

To generate drought-tolerant sugarcane, we obtained hybrid lines by the intergeneric hybridization of a Saccharum spp. hybrid with Erianthus arundinaceus. A test using 5S rDNA as a marker identified five intergeneric hybrid lines with a production efficiency of approximately 3 %. There was difficulty in maintaining these hybrid lines because they were weak, and one line only survived. We examined the amount of roots and polyphenol content of the roots, which are related to drought tolerance, in the surviving line. Although the shoot–root ratio, an indicator of the amount of roots, was high in sugarcane and low in E. arundinaceus, the shoot–root ratio of the hybrid line was intermediate between those of the parents. Regarding the polyphenol content of the roots, only the surface of a root sample was stained by NaNO₃ in sugarcane, whereas the entire cross section of a root sample was stained in E. arundinaceus. For the hybrid line, the entire cross section of the root was faintly stained. From these results, we concluded that the roots characteristics were introduced by the intergeneric hybridization between a Saccharum spp. hybrid and E. arundinaceus.     

Selection of families and parents of sugarcane (Saccharum spp.) through mixed models by joint analysis of two harvests

At present, the sugarcane (Saccharum spp.) breeding programs from around the world have practiced early family selection, because in the early stages of breeding the evaluated characteristics have shown low heritability. This study aimed to select through restricted maximum likelihood/best linear unbiased prediction also known “mixed models” the best families among the 78 that comprise the RB05 sugarcane series, stage T1, of Programa de Melhoramento Genético da Cana-de-açúcar of the Rede Interuniversitária para o Desenvolvimento do Setor Sucroalcoleiro, Brazil. The experiments, conducted during 2007/2008 and 2008/2009 in Paranavaí, PR were implemented using an incomplete-block design with five replicates of each family. Each plot was composed of two planting rows of 5 m, spaced from each other by 1.40 m and spacing between plants was of 0.50 m, containing ten seedlings for planting row, resulting in a total of 20 individuals per replicate of each family. The characteristics evaluated were Brix (°Brix), ton of stalk per hectare (TCH) and ton of °Brix per hectare (TBH). The joint analysis obtained from these three traits in both harvests favored the selection of 35–41 families. The analysis also favored the selection of the best parents. The top five families for Brix were: F41M60, F02M77, F41M82, F61M38 and F62M35; to TCH were F66M30, F35M06, F78M45, F70M30 and F57M46 and for TBH the five elites families were F66M30, F35M06, F78M45, F70M30 and F01M39. The five best sugarcane parents for Brix were RB835486, SP91-1049, SP80-3280, RB72454 and RB925211; to TCH were IAC87-3396, RB941531, RB855511, RB915141 and RB957689, and for TBH the elite were IAC87-3396, RB855511, RB957689, RB941531 and RB855563. As each harvest season had different order of the best families, the joint analysis proved to be a primordial tool for plant breeders.     

Chromosome and cytoplasm analyses of somatic hybrids between onion (Allium cepa L.) and garlic (A. sativum L.)

Chromosomes and cytoplasms were analyzed in two lines of a somatic hybrid between onion (Allium cepa L.) and garlic (A. sativum L.). One line of the somatic hybrid had 40 chromosomes and the other 41chromosomes. Genomic in situhybridization successfully revealed the chromosome constitution of the two lines. One line had 20 chromosomes from onion and17 chromosomes from garlic, and the other had 21 chromosomes from onion and 17chromosomes from garlic. Interestingly, both lines had three chimeric chromosomes. PCR-RFLP analyses of chloroplast and mitochondrial DNAs of both lines showed that these were identical to the onion parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

基于甘蔗热带种(LA-purple)全基因组序列的SSR开发及特征分析

现代甘蔗栽培品种(2n = 100~130)是由甘蔗热带种(2n = 80)与割手密(2n = 40~128)种间杂交而来, 形成异源多倍体、非整倍体作物, 使得甘蔗栽培品种中80%~90%的染色体来源于热带种。开发热带种基因组SSR分子标记, 有助于甘蔗遗传多样性分析、分子标记辅助选择、遗传图谱的构建等。本研究基于热带种LA-purple的全基因组测序数据的255 398个预测基因序列(累计总长为1 029 222 285 bp), 利用Perl程序与生物信息学软件结合, 发掘SSR位点, 获得了153 150个SSR位点, 平均每1.67个基因有1个SSR位点, 其中二、三核苷酸重复基序分别为39 556个和50 072个, 占总SSR位点数的58.5%。在二核苷酸重复基序中, TA/AT所占比例最高, 占41.4%, CG/GC所占比例最低, 占4.6%; 在三核苷酸碱基重复基序中, TGT/ACA所占比例最高, 为15.6%。在TA/AT重复类型中选取100个基序重复次数在60~90之间的SSR位点, 进行引物设计与合成, 在12个甘蔗属材料中进行PCR扩增分析, 从中筛选出52对具有多态性SSR引物, 其中有27对引物在研究的2个甘蔗栽培品种间表现为多态。这些基因组SSR标记的开发, 不仅可以用于甘蔗栽培品种DNA指纹图谱分析, 而且为甘蔗属不同种的遗传图谱构建、遗传多样性分析和重要性状的遗传机制解析奠定基础, 为甘蔗分子育种研究提供重要支撑。     

脱落酸对棉花体细胞胚胎发生的影响

【目的】研究脱落酸(Abscisic acid, ABA)对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24(CCRI 24)下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08μmol·L^-1,分别以A0、A1、A2、A3、A4表示,添加至MSB(MS培养基+B5维生素)培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02μmol·L^-1ABA显著提高胚性愈伤分化率,0.04~0.08μmol·L^-1ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02~0.08μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02μmol·L^-1ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。     

Micropropagation of mother plants of lucerne (Medicago sativa L.) for somatic embryogenesis

Summary A reliable and standard method was established for micropropagation of the A70-34 selected genotype of lecerne (Medicago sativa L., genotype A70-34), aimed at reducing contamination problems, seasonal and phenological influences on regeneration and phytosanitary problems of the mother plants, while maintaining the regenerative potential for somatic embryogenesis of the plant tissues. Mother plants were routinely maintained for several subcultures and somatic embryogenesis was regularly obtained from the subcultured explants. Proliferation, rooting and embryogenetic ability of plants cultured for 30 days was greater than those cultured for 20 days. The regenerative potential of tissues from different organs and of triturated and intact whole plants was also tested. Petioles were confirmed as the best source for embryogenesis as far as efficiency and repeatability were concerned, even though regeneration from other explant types was also achieved. Production of somatic embryos through mechanical trituration of the in vitro cultured plants was obtained; the regeneration ability of the triturated plants was greater than that observed in the intact plants.     

Intertribal somatic hybridization between rapid cycling Brassica oleracea L. and Camelina sativa (L.) Crantz

Black spot, caused by Alternaria brassicae and A. brassicicola, is an important disease in all Brassica oleracea vegetables. Sufficient resistance to the pathogen is not found within the species, nor in species that readily cross to B. oleracea. Camelina sativa (false flax) is highly resistant to Alternaria spp. and has, in addition, other desirable characters for the improvement of B. oleracea. Protoplast fusions were performed between rapid cycling B. oleracea (tribe Brassiceae), which has good regenerability, and C. sativa (tribe Sisymbrieae) by polyethylene glycol (PEG) treatment. The B. oleracea fusion partner was inactivated by treatment with iodoacetate. C. sativa has poor regenerability; hence, no pretreatment was needed for this species. The protoplasts were cultured using a feeder layer system. A total of 2903 calli were isolated from the fusions. Fourteen of these initiated shoots, i.e., 0.5% regeneration frequency. Approximately 110 shoots were excised from 6 of these calli and transferred to rooting medium. Rooted plantlets grew vigorously in vitro and flowering was frequently observed. However, establishment of rooted shoots in soil was unsuccessful. Hybrid identity was confirmed by intermediate shoot morphology, RAPD marker analysis, and flow cytometric estimation of nuclear DNA content. This revised version was published online in August 2006 with corrections to the Cover Date.     

Linkage of genes for anthocyanin pigmentation in rice (Oryza sativa L.)

Summary The inheritance of anthocyanin pigmentation and the interrelationship between genes governing anthocyanin pigmentation in different parts of the rice plant and genes governing panicle density were studied in the cross Suma × Wanar-1. A ratio of 45 pigmented: 19 non pigmented was realised for anthocyanin accumulation in sheath, nodal ring, internode, leaf tip, leaf margin and apiculus whereas a ratio of 3 normal:1 laxy was observed for panicle density. Pleiotropic action of a basic gene (A) and each of the two complementary duplicate genes (P _a or P _b) was detected. A linkage group of genes localising pigment in six plant parts was identified, the sequence of genes being P _nr-P _lm-P _la-P _a-P _sh-P _in. This may form part of the III S _p group of Misro et al. (1966), in which case the genes P _nr and P _la are proposed to be additions to this group.