甘蓝型油菜Ogura 雄性不育×白菜的回交杂种后代与亲本之间蕾期基因表达差异比较研究*

以甘蓝型油菜（Brassica napus L, AACC,2n＝38）Ogura细胞质雄性不育材料为母本,以不同白菜（B campestris ssp. chinensis Makino, AA, 2n＝20） 自交系‘新选一号’和‘矮脚黄’为父本进行杂交,获得了杂种F1、BC1、BC2代。利用cDNA-AFLP技术对两种材料的不同回交世代BC1、BC2代与其亲本在蕾期的基因表达进行分析。结果表明,两种白菜回交世代与其亲本的基因表达有明显差异,在质和量上都存在差异。基因表达模式有5类共7种：（1）单亲沉默型（2种）,（2）单亲一致型（2种）,（3）双亲共沉默型,（4）杂种特异型,（5）表达一致型。随着回交世代的增加,回交杂种和亲本的基因表达在单亲沉默型、双亲共沉默型呈增加趋势。而在母本一致型、父本一致型、杂交种特异型、表达一致型呈下降的趋势。两种白菜在F1、BC1、BC2 3个世代与回交亲本花蕾间的基因差异表达有15种类型,其中以在轮回亲本、F1、BC1、BC2中共同出现表达的带的比例最高。Abstract: Crosses between female parent of Ogura male sterility Brassica napus L. and male parents of B. campestris ssp. chinensis Makino were made and F1, BC1 and BC2 generations produced. Gene expression of two Chinese cabbage backcross hybrid BC1, BC2 and their parents at bud stage was analyzed by means of cDNA-AFLP technique. The results indicated that the patterns of gene expression differ significantly between BC1 and BC2 generations and their parents. There were many patterns of gene expression, including gene overexpression and gene silencing. Five patterns (seven kinds) of gene expression were observed, which include: (1) bands occurring in only one parent (two kinds); (2) bands observed in hybrids and one parent (two kinds); (3) bands occurring in only parents (one kind); (4) bands visualized in only hybrids (one kind); (5) bands observed in parents and hybrids (one kind). In accompany with the addition of backcross, the increase trend in backcross hybrids and their parents were described in the aspects of differential gene expression, bands expressed only in one parent and bands expressed only in both parents. The declined trend in backcross hybrids and their parents were observed in the aspects of bands expressed in both hybrids and one parent (two kinds), bands visualized in only hybrids and bands observed in parents and hybrid. Fifteen patterns of gene expression were observed in F1、BC1、BC2 and backcross parents. The percent of bands expressed in F1、BC1、BC2 and backcross was highest.     

紫孢侧耳、糙皮侧耳及其融合菌株的同工酶分析

采用聚丙烯酰胺凝胶垂直板电泳方法对紫孢侧耳、糙皮侧耳及其融合菌株的酯酶和过氧化物酶同工酶进行了分析。经多次试验,得到了两亲株和融合株恒定的同工酶谱, 在酶谱中融合菌株与亲株有明显差异,并表现出典型的双亲株互补酶带特征,从而证实融合菌株为两亲株基因重组之产物。 Abstract: After many repetitions,the patterns of esterase isozyme and peroxidase isozyme of Pleurotus sapidus,Pleurotus ostreatus and their fusant strains were obtained.There were differences among the parent strains and theirfusant strains in theirisozyme bands.Parental isozyme bands appeared in patterns of fusant strains.Itdemonstrated that fusant were recombinants of P.sapidus and P.ostreatus.     

A representative, highly informative ’genotyping set’ of barley SSRs

We have developed a ’genotyping set’ of 48 SSR-based genetic markers for application in genetical studies of barley. The SSRs are a subset of a collection of approximately 600 SSRs available to the barley research community. They have been specifically chosen according to the following criteria: (1) they are single locus; (2) their product quality is good under standard assay conditions; (3) they are distributed across the barley genome; and (4) they exhibit reasonably high polymorphic information content (PIC) values in the cultivated barley gene-pool. To maximise genotyping throughput, one of each SSR primer pair was 5′ end-labelled with either fam, hex or tet fluorochromes to allow automated data capture after running the samples on a DNA sequencer. SSR product sizes were assembled from a reference set of 24 barley genotypes which allowed the construction of ’graphical genotypes’ of each of the individual lines. The graphical genotypes provide a convenient tool for interrogating genetic similarity in the individuals surveyed. The product sizes were compared to those obtained from end-labelling one of the primers with ³³P and separating the products by denaturing PAGE followed by autoradiography. Although inconsistencies in size were common, they could generally be easily resolved. A reference manual for use of the ’genotyping set’ has been produced and is available as a PDF download file at http://www.scri.sari.ac.uk/ssr/pdf. These well-characterised barley SSRs, for the first time, provide a common set of robust PCR-based tools which can be used to integrate and compare information collected from fundamental and/or applied genetic studies on barley in different laboratories across the world. Received: 17 May 2000 / Accepted: 5 September 2000     

Fitness traits and underlying genetic variation related to host plant specialization in the aphid Sitobion avenae

Sitobion avenae （E） is an important cereal pest worldwide that can survive on various plants in the Poaceae, but divergent selection on different host plants shouldpromote the evolution of specialized genotypes or host races. In order to evaluate their resource use strategies, clones of S. avenae were collected from oat and barley. Host-transfer experiments for these clones were conducted in the laboratory to compare their fitness traits. Our results demonstrated that barley clones had significantly lower fecundityand tended to have longer developmental times when transferred from barley to oat. However, oat clones developed faster after they were transferred to barley. Clones fromoat and barley had diverged to a certain extent in terms of fecundity and developmental time of the nymphs. The separation of barley clones and oat clones of S. avenae was alsoevident in a principal component analysis. Barley clones tended to have higher broad-sense heritabilities for fitness traits than oat clones, indicating the genetic basis of differentiationbetween them. Barley clones showed significantly higher extent of specialization compared to oat clones from two measures of specialization （i.e., Xsp and Ysp）. Therefore, barleyclones were specialized to a certain extent, but oat clones appeared to be generalized. The fitness of S. avenae clones tended to increase with higher extent of specialization. Theevolution toward ecological specialization in S. avenae clones, as well as the underlying genetic basis, was discussed.     

Induction and transmission of wheat-Haynaldia villosa chromosomal translocations

In order to develop more wheat-Haynaldia villosa translocations involving different chromosomes and chromosome segments of H. villosa, T. durum-H, villosa amphiploid was irradiated with ^60Co γ-rays at doses of 800, 1,200, and 1,600 rad. Pollen collected from the spikes 1, 2, and 3 days after irradiation were transferred to emasculated spikes of the common wheat cv. ‘Chinese Spring＇. Genomic in situ hybridization was used to identify wheat-H, villosa chromosome translocations in the M1 generation. Transmission of the identified translocation chromosomes was analyzed in the BC1, BC2, and BC3 generations. The results indicated that all three irradiation doses were highly efficient for inducing wheat-alien translocations without affecting the viability of the M1 seeds. Within the range of 800-1,600 rad, both the efficiency of translocation induction and the frequency of interstitial chromosome breakage-fusion increased as the irradiation dosage increased. A higher translocation induction frequency was observed using pollen collected from the spikes 1 day after irradiation over that of 2 or 3 days after irradiation. More than 70% of the translocations detected in the M1 generation were transmitted to the BC1 through the female gametes. All translocations recovered in the BC1 generation were recovered in the following BC2, and BC3 generations. The transmission ability of different translocation types in different genetic backgrounds showed an order of ‘whole-arm translocation 〉 small alien segment translocation 〉 large alien segment translocation＇, through either male or female gametes, In general, the transmission ability through the female gametes was higher than that through the male gametes. By this approach, 14 translocation lines that involved different H. villosa chromosomes have been identified in the BC3 using EST-STS markers, and eight of them were homozygous.     

Genetic analysis and gene mapping of a rice few-tillering mutant in early backcross populations (Oryza sativa L.)

A rice mutant, G069, characteristic of few tiller numbers, was found in anther culture progeny from the F1 hybrid between an indica-japonica cross, Gui630×02428. The mutant has another two major features: delayed tillering development and yellowing apex and margin on the mature leaves. As a donor parent, G069 was further backcrossed with the recurrent parent, 02428, for two turns to develop a BC2F2 population. Genetic analysis in the BC2F2 population showed that the traits of few-tillering and yellowing apex and margin on the mature leaves were controlled by one recessive gene. A pool of equally mixed genomic DNA, from few-tillering individual plants in BC2F2, was constructed to screen polymorphism with simple sequence repeat (SSR) markers in comparison with the 02428 genome. One SSR marker and three restriction fragment length polymorphism (RFLP) markers were found possibly linked with the recessive gene. By using these markers, the gene of few-tillering was mapped on chromosome 2 between RFLP marker C     

杜仲剥皮再生过程中可溶性蛋白和过氧化物酶含量及分布的变化(英文)

Peroxidases are known to play important roles in plant wound healing. Biochemical analysisand histochemical localization techniques were used to assess changes and distribution of peroxidases inthe recovering bark after girdling in Eucommia ulmoides Oliv. Between 4 and 21 days after girdling (DAG),peroxidases activity in the girdled trees significantly increased by 30-40 times over that in ungirdled trees.During the whole bark recovery process (from 0 to 63 DAG), the peroxidase signal was not found in thetissue regions subjected to intense cell division activity (regenerating cambial zone and phellogen). However,high peroxidase activity was detected in the callus, cortex-like, mature phloem and xylem. Interestingly, itwas shown that, in maturing xylem and phloem cells, there was respectively an inward and outwardperoxidase activity gradient on both sides of the cambium zone. An isoelectric-focusing electrophoresis ofthe extracted protein displayed two isozyme bands of peroxidase: POD Ⅰ and POD Ⅱ. POD Ⅰ was onlydetected in the xylem fraction and could play a role in xylem differentiation. POD Ⅱ was only identified inthe recovering bark portion and could be more engaged in bark regeneration process. A relationshipbetween IAA and peroxidase is also discussed.     

New red flower germplasm lines of cotton selected from hybrid of Gossypium hirsutum × G. bickii

By means of dropping GA3 (50 ppm) and NAA (40 ppm) on the hybrid boll-embryo culture in vitro, one F1 plant of G. hirsutum G. bickii was obtained; when F1 branches were grafted on upland cotton and then back-crossed with upland cotton under short-day and cooler-night condition, some BC1 seeds could be harvested. The characteristic segregation was very violent in early generation. Through 3 times of back-crossing and selecting, ten stable hybrid lines with the character of both male parent (viz. red petal-purple spot and strong fibre) and female par-ent (plant type, earliness, white fibre, lint length, etc. ) were established. These lines were assigned as HB red flow-er lines (HBRL). Transference of character of G. bickii to upland cotton was proved to be successful for the first time. These new germplasms may play an important role in both the genetic research and new cotton variety breeding.     

In vitro selection for Fusarium wilt resistance in Gladiolus

Cormels pieces of four Fusarium susceptible Gladiolus cultivara (Friendship, Peter Pears, Victor Borge and Novalux) formed friable calli when cultured in vitro on Murashige and Skoog basal medium containing various concentrations of auxin and cytokinin. The friable calli established cell suspensions. Plantlet regeneration was obtained from the control callus, control cell suspension derived callus and in vitro selected Fusarium oxysporum Schlecht. resistant cell-lines of Friendship. The in vitro cormlets showed 85-95% germination after breaking dormancy of 8weeks at 4℃. Cell suspensions of all four Gladiolus cultivara were found to be highly sensitive to fusaric acid. Gradual Increase in fusaric acid concentrations to the cell-suspension cultures decreased cell growth considerably. One albino plant was found from the second generation of the In vitro selected cell line of Friendship. The albino plant was found to be highly susceptible to F. oxysporum. The cormlets of all in vitro selected call lines of Friendship were inoculated with a conidial suspension of the F. oxysporum before planting and were also sprayed with the same spore suspension for further characterization when the height of plants was about 6cm. The four selected cell lines showed the same response whether or not they were Inoculated with conidia of the F. oxysporum. Plantlets of all of the selected call lines exhibited significant growth as compared with the control after application of conidia of the F.oxysporum.     

Analysis of QTLs for yield-related traits in Yuanjiang common wild rice (Oryza rufipogon Griff.)

Using an accession of common wild rice (Oryza rufipogon Griff.) collected from Yuanjiang County, Yunnan Province, China, as the donor and an elite cultivar 93-11, widely used in two-line indica hybrid rice production in China, as the recurrent parent, an advanced backcross populations were developed. Through genotyping of 187 SSR markers and investigation of six yield-related traits of two gen- erations (BC4F2 and BC4F4), a total of 26 QTLs were detected by employing single point analysis and inte...     

Establishing a gene trap system mediated by T-DNA(GUS) in rice

Two plasmids, p13GUS and p13GUS2, were constructed to create a gene trap system containing the promoterless β-glucuronidase (GUS) reporter gene in the T-DNA region. Transformation of these two plasmids into the rice variety Zhonghua 11 (Oryza sativa ssp. japonica cv.), mediated by Agrobacterium tumefaciens, resulted in 942 independent transgenic lines. Histochemical GUS assays revealed that 31 To plants had various patterns of the reporter gene expression, including expression in only one tissue, and simultaneously in two or more tissues. Hygromycin-resistant (hygr) homozygotes were screened and the copy number of the T-DNA inserts was determined in the GUS-positivs transgenic plants. The flanking sequences of the T-DNA were isolated by inverse-polymerase chain reaction and the insert positions on the rice genome of T-DNA were determined by a basic local alignment search tool in the GUS-positive transgenic plants transformed with plasmid p13GUS. Moreover, calii induced from the seeds of the T1 generation of 911 GUS-negative transgenic lines were subjected to stress and hormone treatments. Histochemical GUS assays were carried out on the calli before and after treatment. The results revealed that calli from 21 lines displayed differential GUS expression after treatment. All of these data demonstrated that this trap system is suitable for identifying rice genes, including those that are sensitive to induction.     

Comparative study of symmetric and asymmetric somatic hybridization between common wheat and Haynaldia villosa

Symmetric and asymmetric protoplast fusion between long term cell suspension-derived protoplasts of Triticum aestivum (cv. Jinan 177) and protoplasts of Haynaldia villosa prepared from one-year-old embryogeneric calli was performed by PEG method. In asymmetric fusion, donor calli were treated with gamma ray at a dose of 40, 60, 80 Gy (1.3 Gy/min) respectively and then used to isolate protoplasts. Results of morphological, cytological, biochemical (isozyme) and 5S rDNA spacer sequence analysis revealed that we obtained somatic hybrid lines at high frequency from both symmetric and asymmetric fusion. Hybrid plants were recovered from symmetric and low dose γ-fusion combinations. GISH (genomic in situ hybridization) analysis proved exactly the existence of both parental chromosomes and the common occurrence of several kinds of translocation between them in the hybrid clones regenerated from symmetric and asymmetric fusion. And the elimination of donor DNA in hybrid clones regenerated from asymmetric fusion     

暗纹东方TUN 同工酶生化表现型的研究

利用聚丙烯酰胺凝胶垂直平板电泳方法,对暗纹东方|TUN|的心、肝、肾、肌、性腺5种不同组织的7种同工酶(EST、LDH、POD、MDH、SOD、SDH、α-AMY)进行了研究, 讨论了各同工酶的基因表达谱式,观察到EST同工酶存在着多态现象;LDH同工酶有二个基因位点,但只表现3条带,A与B亚基的结合受阻; MDH同工酶存在性别差异,说明决定MDH同工酶表达的因素在不同性别中存在差异;SOD同工酶有3个基因位点。各同工酶酶谱稳定,有组织特异性,但EST、MDH、SOD、POD同工酶在各组织器官中又表现出较大的一致性,有利于物种的鉴定。α-AMY与SDH只在个别组织中有活性,可能与特定组织与器官的形态发生与机能分化有关。 Abstract:By means of polyacrylamid gel electrophoresis seven isozymes(EST.LDH,SOD,MDH,SDH, α-AMY)in heart,liver,kidney,muscle and gonad of Fugu obscurus were studied.The gene expression patterns of each isozyme were analyzed.The results indicated that polymorphism was detected in EST isozymes;LDH isozymes had two loci,but only three bands could be observed,the random association of two subunits(A and B)were restricted;Some MDH isozymes existed sexual differences in identical tissues.The suggested that the factors controlling the expression of MDH isozymes were different between sexes.All the isozyme phenotypes exhibited tissue-specificity and stability,but EST,MDH,SOD and POD isozymes showed relatively consistence in the five tissues.Their characteristic bands could be used in species determination.The activities of SDH andα-AMY isozymes could only be detected in some tissues and closely correlated to the morphological or functional differentiation of those tissues or organs.     

Creation of the technique of interspecific hybridization for breeding in cotton

The new technique of interspecific hybridization was created in Gossypium, which could remarkably overcome abortion of interspecific hybridization and hybrid sterility of F1. A large number of germplasm resources were obtained from seventy cross combinations among the cultivated species and between the cultivated and the 14 wild species, respectively. 8 varieties have been developed, of which 4 were from the cross combination of G. hirsutumXG. arboretum and the other are the first breed from the hybrids between G. hirsutum and 4 wild species, respectively. Of them Shiyuan 321 (jimian 24) is a new variety which had the highest increase in the national Yellow River Valley Regional test, with planting area added up to 933333 ha in the recent three years. The breeding system of interspecific hybridization was established.     

Characterization of transgene integration pattern in F4 hGH-transgenic common carp （Cyprinus carpio L.）

The integration pattern and adjacent host sequences of the inserted pMThGH-transgene in the F4 hGH-transgenic common carp were extensively studied. Here we show that each F4 transgenic fish contained about 200 copies of the pMThGH-transgene and the transgenes were integrated into the host genome generally with concatemers in a head-totail arrangement at 4-5 insertion sites. By using a method of plasmid rescue, four hundred copies of transgenes from two individuals of F4 transgenic fish, A and B, were recovered and clarified into 6 classes. All classes of recovered transgenes contained either complete or partial pMThGH sequences. The class I, which comprised 83% and 84.5% respectively of the recovered transgene copies from fish A and B, had maintained the original configuration, indicating that most transgenes were faithfully inherited during the four generations of reproduction. The other five classes were different from the original configuration in both molecular weight and restriction map, indicating that a few transgenes had undergone mutation, rearrangement or deletion during integration and germline transmission. In the five types of aberrant transgenes, three flanking sequences of the host genome were analyzed. These sequences were common carp β-actin gene, common carp DNA sequences homologous to mouse phosphoglycerate kinase-1 and human epidermal keratin 14, respectively.     

Variation in chromosome number in the seedling progeny of a somaclone of Paspalum dilatatum

The somaclone,C39,derived by tissue culture from the obligate apomict Paspalum dilatatum cv Raki(2n=50),had 50 chromosomes and a karyotype apparently identical to Raki.SC2 seedlings of C39 showed a high degree of phenotypic variation which was often associated with increased chromosome numbers,but some of the variant seedlings were karyotypically indistinguishable from Raki or C39.Plants with increased chromosome numbers exhibited a high degree of intraplant chromosome variation(aneusomaty).In one of the SC2 seedlings,the chromosome number of root tip cells varied from 58 to 82 and in several other seedlings the range was more than 10.The results suggested that the ability to form seed apomictically was much reduced in C39 and that this plant showed some capacity for sexual reproduction and the resulting seedlings,with a chromosome number of about 70,were genetically unstable.Of 11 SC2 seedlings examined cytologically,6 did not produce any viable seed.Seedlings grown from seed of the remaining 5 plants showed that aneusomaty persisted in the SC3 generation.SC3 seedlings which were phenotypically similar to their maternal parent showed a similar range of chromosome numbers to that parent.Some of the SC3 seedlings exhibited an even wider range of chromosome numbers(e.g.56-136),and these plants were all dwarfs.     

Quantifying relationships between rooting traits and water uptake under drought in Mediterranean barley and durum wheat

In Mediterranean regions drought is the major factor limiting spring barley and durum wheat grain yields. This study aimed to compare spring barley and durum wheat root and shoot responses to drought and quantify relationships between root traits and water uptake under terminal drought.One spring barley（Hordeum vulgare L. cv. Rum） and two durum wheat Mediterranean cultivars（Triticum turgidum L. var durum cvs Hourani and Karim） were examined in soil‐column experiments under well watered and drought conditions. Root system architecture traits, water uptake, and plant growth were measured. Barley aerial biomass and grain yields were higher than for durum wheat cultivars in well watered conditions. Drought decreased grain yield more for barley（47%） than durum wheat（30%, Hourani）. Root‐to‐shoot dry matter ratio increased for durum wheat under drought but not for barley, and root weight increased for wheat in response todrought but decreased for barley. The critical root length density（RLD） and root volume density（RVD） for 90% available water capture for wheat were similar to（cv. Hourani） or lower than（cv. Karim） for barley depending on wheat cultivar. For both species, RVD accounted for a slightly higher proportion of phenotypic variation in water uptake under drought than RLD.     

Heterogeneity studies of Carchesium polypinum (Ciliophora: Peritrichida) in Donghu Lake

In this article, RAPD was employed to detect the DNA polymorphism of Carchesium polypinum in three lakes: Nanhu Lake (N), Shahu Lake (S) and Donghu Lake (D). The results were used to deduce the definition of the populations: (1) The 20 primers produced 172 clear bands with an average of 8.6 fragments ranging from 100bp to 1600bp. The other 11 primers produced 59 legible bands with an average of 5.4 fragments ranging from 100bp to 1 500bp; (2) SPSS 12.0 and RAPDistance 1.04 that had similar trees were composed of two parallel main branches. Samples from Shijizhong (Z) of D, Shuiguohu (H) of D and N clustered into a branch and another sample from D — Fengguangcun (F) — gathered with S or made another main branch itself. The result that the three samples from D were separated by the samples from other lakes suggested that the individuals of C. polypinum in D belong to several populations. This discovery will promote micro-hydrobiont population ecology research. In addition, it will help in the study of the essence of the transitional zone and its dispersal and vicariance behavior. We believe that this study will contribute to the study of population differentiation and the mechanism of speciation.     

Loss of Genetic Diversity of Domesticated Panax notoginseng F H Chen as Evidenced by ITS Sequence and AFLP Polymorphism： A Comparative Study with P. stipuleanatus H T Tsai et K M Feng

In the present study, we evaluated the genetic diversity of Panax notoginseng F H Chen, a domesticated species, and P. stipuleanatus H T Tsai et K M Feng, an endangered wild species in southeastern Yunnan and adjacent areas in Vietnam, using sequences of the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA and amplified fragment length polymorphism (AFLP) markers. Twenty-four accessions from three plantations of P. notoginseng and 51 samples from eight populations of P. stipuleanatus were assayed. A total of 694 bp of partial sequences of 18S, ITS 1, 5.8S, ITS2, and partial sequences of 26S were obtained. No sequence variation was detected within P. notoginseng and nine sites (1.30%) were variable in P. stipuleanatus. Two-thirds of the variable sites were found between Langqiao and other populations. In P. notoginseng, four pairs of AFLP primer combinations generated 312 bands, of which 240 (76.9%) were polymorphic and 60.15% of the polymorphisms were harbored within plantations. Approximately 41.0% and 66.9% of bands were polymorphic in population D7 and 5589, respectively. In P.stipuleanatus, the same four primer combinations produced 346 bands, of which 334 (96.5%) were polymorphic and approximately 62.14% of polymorphisms were maintained within populations. Considerable variations were observed. The percentage of polymorphic bands ranged from 50.2% to 84.9% and the average over populations was 70.9%. Cluster analysis did not show correlation of genetic differentiation with the distinctive leaf morphology of P. stipuleanatus (i.e. one form with bipinnatifid leaflets and the other with undivided leaflets). Because over 40% of genetic variations were maintained among populations and because of the very restricted distribution of P. stipuleanatus, all natural populations of this species should be conserved in situ. Considering that there are variations in P. notoginseng within and among plantations, we suggest establishing a genetic resource conservation garden or reintroducing P. notoginseng into its native habitats in southwestern China. Such reintroduction should be carefully executed after large-scale screening of genetic variation within the species.     

Effect of testosterone on long-term organ cultures of canine prostate

Summary Organ cultures of rodent and human prostate glands have shown marked differences in their morphological response to testosterone. In this study, explants from 19 canine prostate glands were cultivated for a minimum of 9 days in Trowell’s T-8 medium. Groups of explants were exposed to media containing from 0.05 to 100 μm testosterone. While the higher testosterone levels (50 and 100 μm) markedly decreased explant viability, explants cultivated at lower levels (0.05 to 5 μm) appeared similar to control explants in testosterone-free Trowell’s T-8 medium. Atmospheric mixtures containing either 95% or 50% oxygen were equally effective. Shortly after the cultures were initiated, large amounts of secretory product were liberated into the lumen. After 9 or more days in vitro, glandular epithelium appeared cuboidal and never revealed the acid phosphatase-rich secretory granules seen in the preculture control. However, the epithelium exhibited an increase in alkaline phosphatase and lipid content following cultivation. This project was supported by contract N01-CP-33331, Carcinogenesis Program, Division of Cancer Cause and Prevention, National Cancer Institute, Bethesda, Maryland.