小麦幼穗胚性愈伤组织诱导及分化过程中内源激素的作用

2,4-D的浓度及诱导时间的长短影响出愈率和早期体胚的形成,6－BA0．25mg／L或KT0．5mg/L抑制胚性愈伤组织的诱导及分化。胚性能力较强外植体的内源激素含量较高,同时分化过程中胚性愈伤组织的ABA、IAA含量高于非胚性意伤组织,而GA3含量则低于非胚性愈伤组织。     

海边香豌豆胚性愈伤组织的诱导和体细胞胚发生

将生长１４ｄ的海边香豌豆（Ｌａｔｈｙｒｕｓ ｍａｒｉｔｉｍｕｓ（Ｌ．）Ｂｉｇｅｌ）无菌苗下胚轴切成０．５ｃｍ左右的片段,置于含有１ｍｇ／Ｌ２,４－Ｄ,０．５ｍｇ／Ｌ ＢＡ和０．５％ＮａＣｌ的ＭＳ培养基中,２８ｄ后诱导出胚性愈伤组织。将其转入含有适当浓度２,４－Ｄ的ＭＳ培养基上,又２８ｄ后可得到大量球形胚和心形胚以及极少量鱼雷胚和子叶胚。诱导体细胞胚适合的２,４－Ｄ浓度为０．５ｍｇ／Ｌ。较高浓度的２     

在离体条件下拐芹(Angelica polymorpha)形成胚性愈伤组织和非胚性愈伤组织过程中几种内源激素的变化(英文)

以拐芹(Angelica potymorpha)的叶柄为外植体,在附加2,4-D(1mg/L) KT(0.5mg/L)MS培养基上诱导脱分化,40天内可形成愈伤组织。在相同的培养基上经过继代培养,可在20天内形成胚性愈伤组织(Embryogenic Callus,EC)和非胚性愈伤组织(Non-Embryogenic Callus,NEC)。将不同天数的培养物作为样品,提取其内源激素;气液相色谱(Gas-Liquid-Chromatography,GLC)测定结果发现,内源ABA,GA_3,CTK和IAA的水平都有变化,尤其是ABA,GA_3。这些结果表明内源ABA和GA_3在胚性愈伤组织和非胚性愈伤组织的分化过程中起着重要作用。     

安祖花胚性愈伤组织诱导及植株再生研究

为构建安祖花(Anthurium andreanum)胚性愈伤组织再生体系,以3个盆栽品种幼嫩叶片和叶柄为外植体,分析了基本培养基、植物生长调节剂组合和培养条件等因素的影响。结果表明,安祖花胚性愈伤组织诱导的最佳培养基为改良MS3+1.5 mg L–1 2,4-D+0.5 mg L–1 KT+4%蔗糖+2%葡萄糖+0.25%Phytagel,且胚性愈伤组织诱导能力差异显著,表现为?粉冠军??罗宾奴??冠军?和叶片叶柄,其中?粉冠军?叶片的胚性愈伤组织诱导率可达57.9%。胚状体分化的最佳培养基为1/2改良MSa+2%蔗糖+0.25%Phytagel,其中?粉冠军?叶片诱导的胚状体分化率可达31.6%,且在光、暗下分化率的差异不显著。分化苗移栽后的成活率可达100%。     

速生湿地松良种胚性愈伤组织诱导与增殖

为使速生湿地松良种快速大规模繁殖,对其胚性愈伤组织进行诱导和增殖优化研究.该文以1代湿地松种子园中10个速生湿地松优良无性系(基因型)的未成熟合子胚为外植体,系统研究基因型、合子胚发育阶段、基本培养基、植物生长调节剂种类和浓度等不同因子对胚性愈伤组织诱导效率的影响,探讨胚性愈伤组织的增殖条件.结果表明:基因型、合子胚发...     

在离体条件下拐芹（Angelica polymorpha）形成胚性愈伤组织和非…

以拐芹（Ａｎｇｅｌｉｃａ ｐｌｏｙｍｏｒｐｈａ）的叶柄为外植体,在附加２,４－Ｄ（１ｍｇ／Ｌ）＋ＫＴ（０．５ｍｇ／Ｌ）ＭＳ培养基上诱导脱分化,４０天内可形成愈伤组织。在相同的培养基上经过继代培养,可在２０天内形成胚性愈伤组织（Ｅｍｂｒｙｏｇｅｎｉｃ Ｃａｌｌｕｓ,ＥＣ）和非胚性愈伤组织（Ｎｏｎ－Ｅｍｂｒｙ－ｏｇｅｎｉｃ Ｃａｌｌｕｓ,ＮＥＣ）。将不同天数的培养物作为样品,提取其内源激素;气液相色谱     

兴安落叶松胚性愈伤组织诱导影响因子的研究

以成熟和未成熟合子胚为外植体,研究影响兴安落叶松（Larix gmelinii）胚性愈伤组织诱导的几种主要因子。结果表明兴安落叶松合子胚带胚乳培养有利于胚性愈伤组织的诱导;内蒙沙地种源成熟合子胚的诱导率显著（p<0.05）高于加格达奇山地种源;冷藏处理可以提高成熟合子胚胚性愈伤组织的诱导率;不同发育时期的未成熟合子胚的诱导率存在显著差别（p<0.05）,其中以子叶初期合子胚（7月5日）诱导率最高;2,4-D对胚性愈伤组织诱导的影响较大,且与BA、KT存在一定的协同作用;S培养基比DCR和MS培养基更有利于胚性愈伤组织的诱导;培养基中琼脂含量为4 g·L^-1时,诱导率较高。     

杉木未成熟胚胚性愈伤组织诱导影响因素探析

该研究从基因型、6-BA浓度、外植体接种方式和合子胚发育阶段等方面,分析杉木未成熟胚胚性愈伤组织诱导的影响因素。结果表明:基因型、6-BA浓度、外植体接种方式和合子胚发育阶段均对胚性愈伤组织诱导频率有不同程度影响。6种基因型中,有3种基因型诱导出胚性愈伤组织,其中基因型S18胚性愈伤组织诱导频率最高,为11.7%。6-BA浓度在1.0~1.5 mg·L~(-1)范围内时,基因型S18的胚性组织诱导频率较高。以在去皮种子的一端切开一个小口的接种方式为最优,将合子胚剥出的方式易造成合子胚褐化死亡,将未剥皮的种子切开一个小口后直接接入培养基的方式不利于愈伤组织生成。适合胚性愈伤组织诱导的合子胚发育阶段为受精至胚器官分化阶段,合子胚进入成熟阶段后不利于胚性愈伤组织诱导,合子胚易生长成完整植株。     

ABA,NAA诱导水稻胚性愈伤组织的研究

ＡＢＡ和ＮＡＡ联合使用能有效地诱导水稻原生质体再生的愈伤组织向胚性发展。通过液体浅层培养由原生质体得到的愈伤性发展。通过液体浅层培养由原生质体得到的愈伤组织,在含ＡＢＡ和ＮＡＡ的Ｎ８培养基上培养一段时间,可以诱导原来呈非胚性状态的愈伤组织形成胚性愈伤组织,并在含ＺＴ的Ｎ６分化培养基上产生绿点。通过对这两种愈伤组织的生化分析,表明二者在游离氨基酸、ＤＮＡ、ＲＮＡ、核酸及蛋白质含量等方面,特别是ＳＤＳ     

Development of microsatellite markers from loquat, Eriobotrya japonica (Thunb.) Lindl

Loquat (Eriobotrya japonica) is a minor fruit which has become an interesting alternative into the European fruit industry. This interest resulted in a loquat germplasm collection established at the Instituto Valenciano de Investigaciones Agrarias, Valencia, Spain. Currently, it is the main reservoir of this species outside Asia. We developed and characterized the first 21 polymorphic microsatellite loci from a CT/AG-enriched loquat genomic library. The observed heterozygosity ranged between 0.20 and 1.00, expected heterozygosity ranged between 0.17 and 0.81, three markers were multilocus and eight loci departed significantly from Hardy-Weinberg equilibrium. These markers will facilitate diversity and genetic studies into the species.     

Hypoglycemic effects of a sesquiterpene glycoside isolated from leaves of loquat (Eriobotrya japonica (Thunb.) Lindl.).

Sesquiterpene glycoside, nerolidol-3-O-alpha-l-rhamnopyranosyl(1-->4)-alpha-l-rhamnopyranosyl(1-->2)-[alpha-l-rhamnopyranosyl(1-->6)]-beta-d-glucopyranoside was isolated from dried leaves of loquat [Eriobotrya japonica (Thunb.) Lindl., Rosaceae]. Hypoglycemic effects of this natural product were assessed in normal and alloxan-diabetic mice model. Animals received orally administration of the sesquiterpene glycoside in dose of 25 and 75 mg/kg. The anti-hyperglycemic effect was compared with gliclazide's. The dose of 25 and 75 mg/kg both exerted a significant (p<0.05) hypoglycemic effect in alloxan-diabetic mice throughout the test and a slight effect in normal mice.     

Genomic characterization of self-incompatibility ribonucleases (S-RNases) in loquat (Eriobotrya japonica Lindl.) (Rosaceae, Pyrinae)

Loquat (Eriobotrya japonica Lindl.) is a fruit tree species of the Pyrinae subtribe of the Rosaceae that behaves as self incompatible. Since self-incompatibility in the Rosaceae is of the gametophytic type where a stylar ribonuclease (S-RNase) controls the female function of pollen–pistil recognition, consensus primers derived from the alignment of S-RNase sequences from other Pyrinae species were used to search for S-RNases in loquat. As a result, the first four S-RNases were sequenced for this species. The genomic sequences obtained showed the structural features of Pyrinae S-RNases. Moreover, microscopic observations of pollen tube growth in the style confirmed the inter-(in)compatibility relationships predicted from the molecular analyses. Phylogenetic analysis of the deduced amino acid sequences with other Pyrinae S-RNases confirmed that divergence of S-alleles in loquat and the Pyrinae predated speciation. This study reports for the first time the genomic characterization of S-RNases in loquat, providing a sound basis for an appropriate selection of pollinator cultivars and an adequate design of breeding programs.     

Isolation of a latent polyphenol oxidase from loquat fruit (Eriobotrya japonica Lindl.): kinetic characterization and comparison with the active form

Polyphenol oxidase (PPO) has been extracted from both soluble and particulate fractions of loquat fruit (Eriobotrya japonica Lindl. cv. Algerie). The soluble PPO (20% of total activity) was partially purified 3.3-fold after ammonium sulfate fractionation being in its active state. The particulate PPO fraction (80% of total activity) was purified to homogeneity in a latent form being activable by sodium dodecyl sulfate (SDS). The enzyme was purified 40.0-fold with a total yield of 15.3% after extraction by phase partitioning in Triton X-114 followed by three chromatographic steps. The molecular weight was estimated to be about 59.2 and 61.2 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, indicating that latent PPO is a monomer. Latent PPO catalyzed the oxidation of chlorogenic acid (CA) at a rate 50-fold faster than that of 4-tert-butylcatechol (TBC) but the soluble active counterpart only twice. Both PPOs exhibited similar Km values for TBC but Km for CA was 5-fold higher for the latent than for the active soluble PPO. Other kinetic characteristics, including sensitivity to inhibitors, substrate specificity, thermal stability, temperature, and pH profiles, were quite different between both PPOs. These results provide strong evidences that the soluble active and the particulate latent are different forms of PPO in loquat fruit flesh. The results suggest that the major PPO form for the oxidation of CA, leading to enzymatic browning under physiological conditions, is the latent one.     

枇杷果肉有机酸组分及有机酸在果实内的分布

用高效离子交换色谱（HPIC）测定了枇杷（Eriobotrya japonica Lindk）18个品种（小毛枇杷、夹脚、卓南1号、解放钟、富阳、森尾早生、华宝2号、香钟10号、白花、土肥、多宝2号、乌躬白、洛阳青、茂木、早钟6号、白梨、塘头4号和长红3号）的成熟果肉和2个品种（解放钟和早钟6号果）成熟果实不同组织有机酸含量。结果表明,成熟果肉中均含有苹果酸、奎尼酸、柠檬酸、异柠檬酸、α-酮戊二酸、富马酸、草酰乙酸、酒石酸8种有机酸,有的还含有微量的阿魏酸、顺乌头酸和B一香豆酸。大多数品种果肉中苹果酸含量最高,平均含量为4399mg kg^-1 FW,占总酸的62．7％;其次是奎尼酸,其平均含量为2042mgkg—FW,占总酸的29．1％。品种之间可滴定酸和有机酸含量差异很大。通过对果肉可滴定酸进行聚类分析,可把18个枇杷品种分为五个组群：极高酸（小毛枇杷）、高酸（夹脚、卓南1号、解放钟和富阳）、中酸（森尾早生、华宝2号、香钟10号、白花、土肥和多宝2号）、低酸（乌躬白、洛阳青、茂木和早钟6号）和极低酸（白梨、塘头4号和长红3号）。解放钟和早钟6号果肉和果皮的总酸含量及可滴定酸均无显著差异,但果皮和果肉的总酸含量和可滴定酸均大大高于种子。相似于果肉,果皮和种子的主要有机酸也是苹果酸和奎尼酸。果皮中苹果酸含量远高于奎尼酸,但种子中苹果酸含量比奎尼酸稍低。此外,种子中苹果酸和奎尼酸比果肉和果皮中的低得多。     

Proteomics of multigenic families from species underrepresented in databases: the case of loquat (Eriobotrya japonica Lindl.) polyphenol oxidases

Here, we approach the problem of obtaining accurate and reliable information about the gene origin of a protein belonging to a multigenic family, polyphenol oxidase, from an underrepresented species, Eriobotrya japonica. De novo sequencing was a key approach to obtain broad sequence coverage. Alignment of peptides on their most similar homologous protein revealed divergent amino acid positions that lead to hypothesize the minimal number of genes encoding for the proteins analyzed.     

Development of two loquat [Eriobotrya japonica (Thunb.) Lindl.] linkage maps based on AFLPs and SSR markers from different Rosaceae species

Loquat [Eriobotrya japonica (Thunb.) Lindl.] is a Rosaceae fruit species of growing interest as an alternative to the main fruit crops. However, only a few genetic studies have been carried out on this species. This paper reports the construction of the first genetic maps of two loquat cultivars based on AFLP and microsatellite markers from Malus, Eriobotrya, Pyrus and Prunus genera. An F₁ population consisting of 81 individuals, derived from the cross between ‘Algerie’ and ‘Zaozhong-6’ cultivars, was used to construct both maps. A total of 111 scorable simple sequence repeat (SSR) loci resulted from the testing of 440 SSR primer pairs in the analyzed progeny and the SSR transferability to Eriobotrya was found to be 74% from apple, 58% from pear and 49% from Prunus spp. In addition, 183 AFLP polymorphic bands were produced using 42 primer combinations. The ‘Algerie’ map was organized in 17 linkage groups covering a distance of 900 cM and comprising 177 loci (83 SSRs and 94 AFLPs) with an average marker distance of 5.1 cM. Self-incompatibility trait was mapped at the distal part of the LG17 linkage group, as previously reported in Malus and Pyrus. The ‘Zaozhong-6’ map covered 870 cM comprising 146 loci (64 SSRs and 82 AFLPs) with an average marker distance of 5.9 cM. The 44 SSRs and the 48 AFLPs share in common by both maps were essentially collinear and, moreover, the order of the 75% of apple and pear SSRs mapped in Eriobotrya was shown to be consistent across the Maloideae subfamily. As a whole, these maps represent a useful tool to facilitate loquat breeding and an interesting framework for map comparison in the Rosaceae.     

箭叶秋葵愈伤组织诱导及增殖培养的激素组合筛选

箭叶秋葵[ Abelmoschus sagittifolius(Kurz.) Merr.]又名红花马宁、铜皮、五指山参、小红芙蓉、岩酸,系锦葵科(Malvaceae)秋葵属(Abelmoschus Medic.)多年生亚灌木状草本;其根既可食用又可入药,具有滋养强壮之功效,可用于治疗神经衰弱、头晕、腰腿痛、胃痛及腹泻等[1].     

Somatic embryogenesis and plant regeneration of litchi (Litchi chinensis Sonn.) from leaves of mature phase trees

Embryogenic cultures were induced from leaflets from new vegetative flushes of mature ‘Brewster’ litchi trees on B5 medium containing 400 mg l⁻¹ glutamine, 200 mg l⁻¹ casein hydrolysate, 30 g l⁻¹ sucrose, 4.52 μM 2,4-D, 9.30 μM kinetin and 3 g l⁻¹ gellan gum in darkness. Embryogenic cultures consisting of proembryonic cells and masses were maintained either on semi-solid MS medium supplemented with 4.52 μM 2,4-D and 0.91 μM zeatin or as embryogenic suspension cultures in liquid medium of the same composition. Maturation of somatic embryos occurred on semi-solid MS medium with 5–20% (v/v) filter-sterilized coconut water in darkness. Recovery of plants from somatic embryos was improved with 14.4 μM GA₃ on half-strength MS medium with 0.2 g l⁻¹ activated charcoal under a 16 h photoperiod provided by cool white fluorescent lights (60–80 μmol s⁻¹ m⁻²). Plants have been successfully acclimatized in the greenhouse.     

An efficient and rapid regeneration via multiple shoot induction from mature seed derived embryogenic and organogenic callus of Indian maize (Zea mays L.)

An efficient method for in vitro micro propagation and genetic transformation of plants are crucial for both basic and applied research. Maize is one of the most important cereal crops around the world. Regeneration from immature embryo is hampered due to its unavailability round the year. On the contrary mature embryo especially tropical maize is recalcitrant toward tissue culture. Here we report a highly efficient regeneration (90%) system for maize by using 2 different approaches i.e., embryogenic and organogenic callus cultures. Seeds were germinated on MS medium supplemented with 5 mg/l 2,4-D and 3 mg/l BAP. Nodal regions of 2 wks old seedlings were longitudinally split upon isolation and subsequently placed on callus initiation medium. The maximum frequency of embryogenic callus formation (90%) was obtained on MS medium supplemented with 2 mg/l 2,4-D and 1 mg/l BAP in the dark conditions. The compact granular organogenic callus formation (85% frequency) was obtained on MS medium supplemented with 2.5 mg/l 2,4-D and 1.5 mg/l BAP at light conditions. MS medium supplemented with 2 mg/l BAP, 1 mg/l Kinetin and 0.5 mg/l NAA promoted the highest frequency of shoot induction. The highest frequency of root formation was observed when shoots were grown on MS medium. The regenerated plants were successfully hardened in earthen pots after adequate acclimatization. The important advantage of this improved method is shortening of regeneration time by providing an efficient and rapid regeneration tool for obtaining more stable transformants from mature seeds of Indian tropical maize cultivar (HQPM-1).