栽培番茄×秘鲁番茄杂交后的胚胎发育

“北京早红”番茄(Lycopersicon esculentum)×秘鲁番茄(L.peruvianum)PI,128657中的8号株系杂交后杂种种子不能正常发育的原因观察。结果表明,杂种胚和胚乳发育缓慢。授粉后16天胚乳开始退化。杂种胚的分裂是随机的,没有经历正常的发育阶段。授粉后30天杂种胚开始退化,35天完全解体。授粉后4天有些珠被绒毡层出现增生。     

秘鲁番茄和多毛番茄的组织培养及植株再生

1植物名称秘鲁番茄（Lycopersiconperuviaman）、多毛番茄（L．hirsutum）。2材料类别真叶。3培养条件种子播种于珍珠岩和菜园土（1：1）的混合培养上中。子叶展开后每隔1周浇of％尿素。取第3片完全展开的真叶作外植体。基本培养基为MS,添加3％蔗糖、07％琼脂,PH58。（1）愈伤组织诱导培养基：MS＋6－BA2mg·L’（单位下同）＋IAAO4（秘鲁番茄）;MS十个BA2＋NAA几2（多毛番茄）。（2）芽分化培养基：秘鲁番茄和多毛番茄均为MS＋6BAZ。（3）生根培养基：MS－NAA0．l。培养温度25士Ic,照光16h·d－‘,光照度2000IX。4生长…     

番茄基质通气栽培模式的效果

针对雾培模式在提高作物产量同时增加无土栽培成本的问题,研制了一种新型的珍珠岩通气栽培模式,探讨了其对番茄的栽培效果.试验设计3种栽培方式:全珍珠岩栽培(CK),珍珠岩通气栽培(T1)和气雾培(T2).结果表明:T1可显著改善番茄根际通气环境,其中根际CO2浓度仅为CK的1/5,O2浓度则为CK的1.17倍;显著增加了番茄的株高和茎粗,在定植后60d时,株高和茎粗分别比CK增加了5.1%和8.4%;植株净光合速率显著高于CK,在净光合速率达到最大值(定植后45d)时,比CK提高了13%;显著提高了植株根系活力和吸收能力,在定植后45d时,其根系活力为CK的1.23倍,在定植后60d时,根系钾、钙、镁含量分别比CK增加了31%、37%和27%,番茄产量为CK的1.16倍.且T1上述指标均与T2无显著差异;而CK、T1和T2在果实的可溶性糖、有机酸、糖酸比方面无显著差异.表明以珍珠岩为基质的通气栽培模式简便易行且可显著提高番茄产量.     

温室有机土栽培CO2浓度变化规律及增施CO2对番茄生长发育的影响

实验分析了有机土栽培下温室内CO2浓度变化规律,研究了增施CO2对温室番茄植株生理效应、产量、果实品质的影响.结果表明有机土栽培条件下温室内CO2浓度变化存在明显的季节变化和日变化.温室内CO2浓度在11月和3月,最高浓度达到1 200 μL·L-1以上,在改善温光条件下,可不施或少施CO2;而7月温室内CO2日最高浓度在500 μL·L-1以下,每天应提早增施CO2.CO2空间分布为近地面层》畦面》植株内部》冠层》株顶上部.不同的栽培方式下,有机土壤栽培CO2浓度日变化范围为331～1 294 μL·L-1,而外界浓度与土壤无作物栽培方式日变化范围为327～556 μL·L-1,土壤栽培CO2变化范围为402～1 047 μL·L-1.光照强度是影响温室内CO2浓度和利用效率的主要因素.与对照相比,温室内增施CO2番茄株高增加18.29%,总干重增加18.69%,功能叶面积增加22.02%,光合速率提高48.92%,叶绿素含量增加33.00%,羧化效率提高87.50%,产量增加 26.48%,果实Vc增加33.27%,番茄红素增加30.98%,差异均达到显著水平.     

番茄属种间杂种离体培养F1植株的细胞学分析

本文报道了栽培番茄（Lycopersico esculentum)“北京早红”等5个品种分别与野生型秘鲁番茄（L.peruvianum)PI128657中8号株系杂交,离体胚培养,得F1杂种植株,对花粉母细胞在减数分裂中染色体行为和终变期二价体交叉点的频率,以及亲和性程度等进行了.结果表明,6个亲本植株花粉母细胞减数分裂染色体的行为是正常的,中期I为12个二价体。其中环状二价体占多数,棒状二价体数较少,中期I没有单价体,后期I和II均正常,四分体阶段无微出现,但各亲本在终变期和中期I的环状二价体和棒状二价体的数有一定的差异,这可能与不同亲本基因型的亲和性程度和在遗传学上的不协调有关。5个组合的大多数F1杂种花粉母细胞减数分裂中染色体行为基本正常,12个二价体占多数,但染色体配对不稳定,有较多的单价体,染色单体桥。四分体阶段有微核,此外,在5个组合的F1杂种植株中,均出现双二倍体花粉母细胞,这些双二倍体花粉母细胞的染色体,在减数分裂中,也均,出现落后染色体和染色单体桥,以及较多的多价体,四分体阶段有微和不同类型的四分孢子群。     

不同倍性香石竹杂交受精过程及胚胎发育研究

采用石蜡切片法对以四倍体香石竹品种‘紫蝴蝶’(2n=4x=60)为母本,单瓣中间材料‘NH6’(2n=2x=30)为父本杂交后受精过程及胚胎发育进行研究。结果表明:(1)授粉后17h,花粉管进入助细胞并释放内容物,精核进入极核细胞内,与二极核细胞融合形成初生胚乳核;授粉后1d,精核向卵核方向移动,贴伏于卵核核膜上;授粉后2d,形成合子及游离的胚乳核;随后,胚发育经过原胚、球形胚、心形胚、鱼雷形胚阶段。(2)杂交障碍发生在受精过程及胚胎发育的各个时期,表现为:精子与卵细胞不相融合或精子与二极核不相融合、合子未分裂或初生胚乳核未分裂及胚胎的败育。(3)胚败育虽能发生在原胚、球形胚、棒状形胚、三角形胚、心形胚、鱼雷形胚及子叶形胚阶段,但主要发生在球形胚阶段。     

侵染番茄的番茄花叶病毒的研究

从种传番茄苗中获得一病毒分离物Ｔｏ－Ｓｌ,人工摩擦接种７科２４种植物,Ｔｏ～Ｓｌ能侵染４科１５种植物,在番茄上产生花叶,在白肋烟上为局部枯斑。Ｔｏ－Ｓｌ的钝化温度为８５～９０℃,稀释限点为１０￣（－６）～１０￣（－７）．体外保毒期在一个月以上。病毒粒体杆状,长度主要分布于２８１～３００ｎｍ之间,平均长度２８８ｎｍ。病毒衣壳蛋白亚基只有一条多肽链,分子量为２１ｋＤａ。ｄｓＲＮＡ分析测得其基因组长度为６．４ｋｂ。琼胎糖双扩散和胶内交叉吸附试验证明,Ｔｏ－Ｓｌ与ＴＭＶ有血清关系,但有一定的差异,病毒粒体电泳分析也表明Ｔｏ－Ｓｌ与ＴＭＶ粒体有差异。在交叉保护试验中,ＴＭＶ和Ｔｏ－Ｓｌ之间均无保护作用。根据以上试验结果Ｔｏ－Ｓｌ被鉴定为番茄花叶病毒。这是我国首次系统报道番茄上番茄花叶病毒的侵染。     

普通番茄四倍体与二倍体杂交的杂种不育性研究

以普通番茄的四倍体为母本与二倍体杂交时,花粉萌发、花粉管生长及双受精过程都正常;对受精的胚珠形态结构观察得知:4天前幼胚和胚乳发育正常,授粉后4天胚乳出现异常开始退化,5天时这种异常现象更明显,第7天胚乳已完全解体。幼胚在胚乳完全解体前发育正常,而胚乳解体后,幼胚也开始解体。所以普通番茄二倍体与四倍体杂交的杂种不育的原因,在于胚乳败育以及胚的死亡。     

滴灌频次对温室长季节栽培番茄产量和养分利用效率的影响

为提高日光温室长季节栽培番茄的养分和水分利用效率,明确其最佳滴灌施肥制度,本研究以番茄嫁接苗为试材,采用土壤栽培和水肥一体化膜下滴灌方式开展试验。为计算养分利用效率,以常规水溶肥(平衡肥N-P₂O₅-K₂O=20-20-20和高钾肥N-P₂O₅-K₂O=17-8-30)每12 d滴灌1次作为对照(CK),以每12 d滴灌1次清水作为CK₁,处理组(T₁～T₄)均滴灌日本山崎(1978)番茄配方营养液,在确保每12 d灌水施肥总量一致的前提下,分别采用每2 d(T₁)、4 d(T₂)、6 d(T₃)和12 d(T₄)滴灌1次的频次进行,研究滴灌频次对长季节栽培番茄产量和养分利用效率的影响。结果表明：随滴灌频次减少,番茄的产量、干物质和氮磷钾积累量、肥料偏生产力和养分利用效率均呈先升高后降低趋势,在T     

Asymmetric somatic hybrids between Lycopersicon esculentum and irradiated Lycopersicon peruvianum

Summary Asymmetric somatic hybrids of Lycopersicon esculentum and Lycopersicon peruvianum were analysed for the retention of genes and alleles specific for L. peruvianum. The hybrids were obtained by fusion of protoplasts from L. esculentum with those of L. peruvianum (the donor), the latter having been irradiated before fusion with 50, 300 or 1,000 Gy of gamma-rays. The retention of three different types of genes or alleles was analysed. (1) The gene coding for kanamycin resistance, which is dominant and had been introduced in most of the L. peruvianum donor plants by transformation. It was present at one locus in 16 L. peruvianum donor plants and at two loci in one donor plant. (2) The genes coding for acid phosphatase, locus Aps-1, and glutamate oxaloacetate transaminase (GOT); different alleles of these genes are co-dominant and were detected by isozyme analysis. (3) Eighteen single gene morphological markers for which most of the L. esculentum genotypes used were homozygous recessive. Kanamycin resistance from donor plants with one locus was retained in about 50% of the asymmetric 30H-hybrids (the donor was irradiated with 300 Gy). L. peruvianum specific alleles of Aps-1 and GOT were present in at least 70% of the hybrids; the retention of donor alleles was lower in 30H- than in 5H-hybrids (donor irradiated with 50 Gy). On average, 73% of the L. peruvianum-specific alleles (one or both) of the morphological markers were detected in the 30H-hybrids. Several of the L. esculentum genotypes used were homozygous recessive for two morphological markers on the same chromosome; in 43% of the 30H-hybrids derived from them, only one of these markers was complemented by the L. peruvianum allele. This is an indication of frequent breakage of the L. peruvianum chromosomes. Several hybrid calli regenerated genotypically different shoots. On the whole, this analyses confirms the conclusion drawn from the cytogenetic and morphological analysis of these asymmetric hybrids, namely that irradiation prior to fusion eliminates the L. peruvianum genome to only a limited extent.     

Asymmetric somatic hybrids between Lycopersicon esculentum and irradiated Lycopersicon peruvianum

Summary Asymmetric somatic hybrids of Lycopersicon esculentum and Lycopersicon peruvianum were obtained by fusion of leaf protoplasts from both species after irradiation of protoplasts or leaf tissue of L. peruvianum with 50, 300, or 1,000 Gy of gamma-rays. These radiation doses were sufficient to abolish the growth of the L. peruvianum protoplasts. The hybrids were selected for their ability to regenerate plants; this regeneration capacity derived from L. peruvianum. All asymmetric hybrid plants were aneuploid. The ploidy level, the morphology, and the regeneration rate were analyzed in relation to the radiation dose applied to L. peruvianum. After a low dose (50 Gy), most hybrids had near-triploid chromosome numbers, whereas after a high dose (300 or 1,000 Gy), most hybrids had near-pentaploid numbers. The morphology of the asymmetric hybrids was intermediate between that of L. esculentum and symmetric somatic hybrids of both species (obtained without irradiation treatment), and approached the morphology of L. esculentum to a greater extent after a high dose of irradiation. The asymmetric hybrids regenerated more slowly than the symmetric hybrids and regeneration proceeded more slowly after a high dose than after a low dose of irradiation. The high-dose hybrids also grew more slowly, flowered less, and set fruits less than the low-dose hybrids. No seeds could be obtained from any asymmetric hybrid.     

Organelle analysis of symmetric and asymmetric hybrids between Lycopersicon peruvianum and Lycopersicon esculentum

Summary The organelles of somatic hybrids obtained from symmetric and asymmetric fusions between the Lycopersicon species L. peruvianum and L. esculentum were analyzed by DNA hybridization methods. In the asymmetric fusions the L. peruvianum protoplasts were gamma-irradiated at a dose of 50, 300 and 1,000 Gy. The organelles were characterized using the Petunia chloroplast probe pPCY64 and the mitochondrial EcoRI-SalI fragment of the Pcf gene. In all symmetric and asymmetric hybrid plants, a total of 73 being analyzed, only one of the parental chloroplast genomes was present, except for one hybrid plant which harbored both parental chloroplast genomes. No recombination and/or rearrangement in the chloroplast genome could be identified with the pPCY64 probe. Irradiation of the L. peruvianum protoplasts did not significantly reduce the fraction of asymmetric hybrids with L. peruvianum chloroplasts. A novel mitochondrial restriction pattern was present in 5 out of 24 hybrids tested. In 9 hybrids novel combinations of chloroplasts and mitochondria were found, indicating that both organelle types sorted out independently.     

The development of bridge lines for interspecific gene transfer between Lycopersicon esculentum and L. peruvianum

Summary Using a modified embryo callus culture technique, hybrids between Lycopersicon esculentum and L. peruvianum were developed and their usefulness as bridge lines for facilitating interspecific gene transfer was evaluated. Four of these lines showed a high level of sexual compatibility with several other L. peruvianum var. typicum accessions, as well as with accessions of L. peruvianum var. humifusum and L. peruvianum var. glandulosum and L. esculentum. These bridge line x L. peruvianum hybrids could be crossed with L. esculentum to introgress genes from L. peruvianum into L. esculentum.     

Clonal propagation in vitro from immature embryos and flower buds of Lycopersicon peruvianum and L. esculentum

Direct propagation of Lycopersicon esculentum Mill. and L. peruvianum (L.) Mill. has been achieved from both reproductive and embryonic stages of the life cycle. Multiple adventitious shoots were induced from immature flower buds on Murashige and Skong (MS) basal medium supplemented with 2 mg 1⁻¹ 6-benzylaminopurine (BAP). Multiple shoots and somatic embryos were induced from immature zygotic embryos on HLH basal medium supplemented with 1 g 1⁻¹ yeast extract and 2 mg 1⁻¹ BAP. The nature of the response was related to the developmental stage of the parent embryo at explanting. The process of multiple shoot induction from embryos shows features in common with direct somatic embryogenesis from sexual embryos of Lycopersicon and other genera.     

Pachytene pairing and metaphase I configurations in a tetraploid somatic Lycopersicon esculentum x L. peruvianum hybrid.

In the tetraploid somatic hybrid between the diploid Lycopersicon species L. esculentum (tomato) and L. peruvianum, synaptonemal complexes formed quadrivalents in 73 of the 120 sets of four chromosomes (60.8%) in 10 cells studied in detail at pachytene. Of these, 43 had one pairing partner exchange, 22 had two, and 8 had three, very close to a Poisson distribution. The points of pairing partner exchange were concentrated at the middle of the two arms. The frequency per arm corresponded with physical arm length. There was a sharp drop around the centromere, and pericentric heterochromatin had a slightly lower probability of being involved in pairing partner exchange than euchromatin. The chromosomes align before pairing and there are several points of pairing initiation, with concentrations at or near the ends and the centromere. From zygotene to late pachytene the quadrivalent frequency decreased considerably. At late pachytene it was lower than expected with the observed high frequency of pairing partner exchange. Pairing affinity between species was only slightly lower than affinity within species, in spite of considerable genetic differentiation. The frequency of recombination nodules increased from early to late zygotene and then decreased strongly to full pachytene. There is a highly significant negative correlation between percent pairing and SC length. At metaphase I the frequency of quadrivalents was 0.444, and branched quadrivalents were rare, probably caused by interference and restriction of chiasma formation to distal euchromatin. Metaphase I quadrivalent frequency is a relatively good indication of pairing affinity in this material.     

Homoeologous pairing and recombination in backcross derivatives of tomato somatic hybrids [Lycopersicon esculentum (+) L. peruvianum]

 Genomic in situ hybridization (GISH) was used to examine genome interactions in two allohexa ploid (2n=6x=72) Lycopersicon esculentum (+) L. peruvianum somatic hybrids and their seed progenies originated from subsequent backcrosses to L. esculentum. The ability of GISH to distinguish between chromatin derived from two closely related species, L. esculentum and L. peruvianum (both 2n=2x=24), allowed the precise chromosomal constitution of somatic hybrids and their backcross progenies to be unequivocally established. This enabled the interaction of species genomes to be observed at meiosis, providing clear evidence of strictly regular homoeologous pairing and the high degree of homoeologous recombination in allodiploid plants (2n=2x=24) of the BC₁ generation. In segmental allodiploids of the BC₂ and BC₃ generations, the recombinant chromosomes continued to pair with a homoeologous partner (in the absence of a homologous one), and therefore could be stably incorporated into gametes. Chiasmata were found almost exclusively in more distal, rather subterminal, chromosome segments. A considerable proportion of meiotic recombination was detected in subterminal heterochromatic regions, often involving distal euchromatin, located in close proximity. GISH also supplied information on the extent of the overall sequence homology between the genomes of L. esculentum and L. peruvianum, indicating that despite their different breeding systems, these species may not be differentiated to a high degree genetically. The present study has demonstrated that somatic hybridization between two such closely related, but sexually incompatible or difficult to cross species, provides a way of transferring genes, via homoloeogous crossing-over and recombination, across the incompatibility barriers. Indeed, such hybrids may offer the preferred route for gene transfer, which subsequently results in more stable gene introgression than other methods. Received: 22 July 1996 / Accepted: 23 August 1996     

Purification of extensin from cell walls of tomato (hybrid of Lycopersicon esculentum and L. peruvianum) cells in suspension culture

Extensin, a hydroxyproline-rich glycoprotein comprising substantial amounts of -l-arabinose-hydroxyproline glycosidic linkages is believed to be insolubilized in the cell wall during host-pathogen interaction by a peroxidase/hydroperoxide-mediated cross-linking process. Both extensin precursor and extensin peroxidase were ionically eluted from intact water-washed tomato (hybrid) of Lycopersicon esculentum Mill. and L. peruvianum L. (Mill.) cells in suspension cultures and purified to homogeneity by a rapid and simple procedure under mild and non-destructive experimental conditions. The molecular weight of native extensin precursor was estimated to be greater than 240–300 kDa by Superose-12 gel-filtration chromatography. Extensin monomers have previously been designated a molecular weight of approximately 80 kDa. Our results indicate that salt-eluted extensin precursor is not monomeric. Agarose-gel electrophoresis, Superose-12-gel-filtration, extensin-peroxidase-catalysed cross-linking, Mono-S ion-exchange fast protein liquid chromatography (FPLC), and peptide-sequencing data confirmed the homogeneity of the extensin preparation. Evidence that the purified protein was extensin is attributed to the presence of the putative sequence motif — Ser (Hyp)₄ — within the N-terminal end of the protein. Treatment of extensin with trifluoroacetic acid demonstrated that arabinose was the principal carbohydrate. The amino-acid composition of the purified extensin was similar to those reported in the literature. The cross-linking of extensin in vitro upon incubation with extensin peroxidase and exogenous H₂O₂ was characteristic of other reported extensins. Furthermore, Mono-S ion-exchange FPLC of native extensin precursor resolved it into two isoforms, A (90%) and B (10%). The amino-acid compositions of extensin A and extensin B were found to be similar to each other and both extensins were cross-linked in vitro by extensin peroxidase.Abbreviations CM-cellulose carboxymethyl-cellulose - FPLC fast protein liquid chromatography - HF hydrogen fluoride - HRGP hydroxyproline-rich glycoprotein - Hyp hydroxyproline - V_c retention volume - TCA trichloroacetic acid - TFA tri-fluoroacetic acid This work was supported by a A.F.R.C. postdoctoral assistantship to Michael D. Brownleader. We thank Dr. Anthony K. Allen (Department of Biochemistry, Charing Cross and Westminster Hospital, London, UK) for performing the amino-acid analysis and Mrs. Margaret Pickering (Department of Biochemistry, Royal Holloway) for performing the peptide-sequence analysis of extensin. We also express our gratitide to Dr. A. Mort (Oklahoma State University) for performing the HF-deglycosylation of extensin.