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[Determination of the true digestibility of amino acids in variously prepared potatoes in growing pigs. 1. Freshly steamed potatoes and silage made from steamed potatoes].

Freshly Steamed Potatoes of 6 varieties were investigated. With 3 varieties the true N and amino acid digestibilities were found to be in good agreement, while great variations were found in potatoes of the other 3 varities so that it appears to be a matter of some doubt whether the true amino acid digestibility could, in the case of potatoes, be reliably derived from data on the true N digestibility. Differences in the true amino acid digestibility data were found between the varities of potatoes grown in different years. A positive trend was found to exist in the relationship between the absolute amino acid content and true digestibility. The true lysine digestibility of freshly steamed potatoes varies between 76-90%, the average percentage being about 80%. The true methionine digestibility was found to vary within still wider limits, although the average percentages was also about 80%. Values established for the true cystine digestibility ranged from 80% to 90%. The true amino acid digestibility data did not provide a sufficiently reliable basis for making conclusions as to the existence of a limiting amino acid. Both the lowest value for true digestibility and the greatest variations were found in the case of the amino acids alanine, tyrosine, and methionine. It was observed that silage made from steamed potatoes had a lower true digestibility of N than steamed fresh potatoes although the value for true amino acid digestibility were the same, with the exception of methionine and cystine.

Amino Acids

Engineering resistance to mixed virus infection in a commercial potato cultivar: resistance to potato virus X and potato virus Y in transgenic Russet Burbank.

Potato virus X (PVX) and potato virus Y (PVY) infection in potato may result in the loss of certification of seed potatoes and affect quality and yield of potatoes in commercial production. We transformed a major commercial cultivar of potato, Russet Burbank, with the coat protein genes of PVX and PVY. Transgenic plants that expressed both CP genes were resistant to infection by PVX and PVY by mechanical inoculation. One line was also resistant when PVY was inoculated with viruliferous green peach aphids. These experiments demonstrate that CP protection is effective against mixed infection by two different viruses and against mechanical and aphid transmission of PVY.

Amino Acid Sequence

Expression pattern of Stlhcb gene family in potato and effects of overexpression of Stcp24 gene on potato photosynthesis.

Potato is one of the four staple food crops in the world. It has a wide range of cultivation, high yield, and high nutritional value. Enhancing the photosynthesis of potato is particularly important as it leads to an increase in the potato yield. The light-harvesting pigment-binding protein complex is very important for plant photosynthesis. We identified 12 Stlhcb gene family members from the potato variety "Atlantic" using transcriptome sequencing and bioinformatics. The proteins encoded by the Stlhcb gene family have between 3358 and 4852 atomic number, a relative molecular weight between 24060.16 and 34624.54 Da, and an isoelectric point between 4.99 and 8.65. The RT-qPCR results showed that the 12 Stlhcb genes were expressed in a tissue-specific and time-dependent fashion under low light. The relative expression of the Stlhcb genes in the leaves was significantly higher than that in the stems and roots, and the relative expression of these genes first increased and then decreased with the prolongation of light exposure time. The Stcp24 gene with the highest expression was cloned, and an expression vector was constructed. A subcellular localization analysis was performed in tobacco and an overexpression experiment was performed in potato using an Agrobacterium-mediated method. The subcellular localization analysis showed that the protein encoded by Stcp24 was located in chloroplasts as expected. Overexpression of Stcp24 in transgenic potato increased the yield of potatoes and the content of chlorophyll a and b; increased the net photosynthetic rate, transpiration rate, stomatal conductance, electron transport efficiency, and semi-saturated light intensity; and promoted photosynthesis and plant growth. This study provides a reference for the study of the function of the potato light-harvesting pigment-binding protein gene family. It lays a foundation for further study of the mechanism of the photosynthesis of potato, improvement of the light energy utilization of potato, and molecular breeding of potato.

Solanum tuberosum

[Studies on growing pigs to determine the true digestibility of variously prepared potatoes. 2. Technically dried potatoes].

There are two kinds of damage to potato proteins that may arise in the process of beat drying. Decreases in the gross lysine content are accompanied by a still much more pronounced depression of digestibility. If, for example, the gross lysine content of dried potatoes was only 50% of that of steamed potatoes then the average value for the true lysine digestibility was 15%; in some cases the content of truly digestible lysine was 0% when compared to that of steamed potatoes. Similarly, it was found that some types of dried potatoes with a dry matter content of about 88% had already lost about 43% of the truly digestible lysine contained in steamed potatoes.

Animal Nutritional Physiological Phenomena

The R1 gene conferring race-specific resistance to Phytophthora infestans in potato is located on potato chromosome V.

Late blight in potato is caused by the fungus Phytophthora infestans and can inflict severe damage on the potato crop. Resistance to P. infestans is either based on major dominant R genes conferring vertical, race-specific resistance or on "minor" genes inducing horizontal, unspecific resistance. A dihaploid potato line was identified which carried the R1 gene, conferring vertical resistance to all P. infestans races, with the exception of those homozygous for the recessive virulence allele of the locus V1. The F1 progeny of a cross between this resistant parent P(R1) and P(r), a line susceptible to all races, was analysed for segregation of R1 and of restriction fragment length polymorphism (RFLP) markers distributed on the potato RFLP map comprising more than 300 loci. The R1 locus was mapped to chromosome V in the interval between RFLP markers GP21 and GP179. The map position of R1 was found to be very similar to the one of Rx2, a dominant locus inducing extreme resistance to potato virus X.

Chromosomes

Multiple forms of soluble monophenol, dihydroxyphenylalanine: oxygen oxidoreductase (EC 1.14.18.1) from potato tubers (Solanum tuberosum). III. Influence of pH on the molecular weight distribution of enzyme activity in potato juice.

Gel chromatography on Sepharose and on Sephadex was used to separate the soluble phenol oxidase in various potato juices into multiple molecular forms ranging from 36,000 to 800,000 daltons. Adjustment of potato juice from physiological pH (ca. 6) to pH 4.5 or to pH 7.8 resulted in the predominance of low-mol.-wt. (less than 150,000 daltons) or high-mol.-wt. (greater than 150,000 daltons) enzyme forms, respectively. This suggests association phenomena of subunits. In potato juice of physiological pH and in potato juice adjusted to pH 4.5, all enzyme forms exhibited both monophenol and o-diphenol oxidase activities (assayed at pH 6.0). In potato juice adjusted to pH 7.8 considerable loss of monophenol oxidase activity (assayed at pH 6.0) occurred. This suggests that o-diphenol oxidase is more alkali-stable than monophenol oxidase. The significance of these findings for enzyme purifications and for the in vivo action of the enzyme is discussed.

Catechol Oxidase

Expression of the potato leafroll luteovirus coat protein gene in transgenic potato plants inhibits viral infection.

Transgenic potato plants, cultivar Désirée, were produced that contained the coat protein gene of potato leafroll luteovirus (PLRV). The transformed potato plants expressed the PLRV coat protein (CP) RNA sequences but accumulation of coat protein in transgenic tissues could not be detected. Upon inoculation with PLRV, the PLRV CP RNA expressing potato plants showed a reduced rate of virus multiplication.

Base Sequence

Resistance to phloem transport of potato leafroll virus in potato plants.

A 'double-graft sandwich' technique in which sections of potato stem from different potato cultivars were grafted between a susceptible healthy stock plant and a potato leafroll virus (PLRV)-infected scion was used to study the rate of phloem transport of PLRV in cultivars differing in resistance to PLRV infection (IR) and accumulation (AR). Resistance to phloem transport (i.e. delayed PLRV systemic movement) was found in Bismark cultivar (IR A(S)). This was independent of IR and AR as the rate of movement in Bismark cultivar was markedly slower than that in Omega and Spunta (IR AR), Delaware (I(S) AR), and Desiree and Renova (I(S) A(S)) cultivars. It operated in Bismark cultivar stems of two different ages, but did not operate against potato virus X (PVX) and was not influenced by previous infection with this virus. Aphid vector (Myzus persicae) feeding preferences and colonization rates differed between cultivars, but the cultivar characteristics responsible were unrelated to IR, AR or resistance to phloem transport. Delayed systemic movement of PLRV out of leaves inoculated with viruliferous aphids was independent of AR and resistance to phloem transport, and remained unaffected by previous infection with PVX. It was also independent of cultivar factors causing different aphid feeding preferences and colonization rates, but may be linked to IR.

Biological Transport

Increased resistance to potato virus X and preservation of cultivar properties in transgenic potato under field conditions.

During the last three years we performed field trials to assess levels of resistance against potato virus X (PVX) and changes in intrinsic properties of the potato cultivars Bintje and Escort upon the introduction of the PVX coat protein (CP) gene. Analysis of leaf and tuber samples collected in the field at two week intervals revealed a stable expression of the PVX CP gene throughout the growing season. This resulted in a large decrease in PVX incidence among clonal progeny obtained from previously infected Bintje and Escort clones. Based on evaluation of 50 defined morphological characteristics, tuber yield and grading, 81.8% of the Escort and 17.9% of the Bintje derived transgenic clones proved to be true to type. Overall lightsprout morphology was a useful criterion for the early detection of deviant transgenic clones. Using the polymerase chain reaction (PCR) with convergent primers spanning transgenic sequences, true to type clones could be distinguished unambiguously from the corresponding untransformed cultivars. Clear distinctions between independent transgenic clones could be made by inverted PCR (IPCR) diagnosis revealing integration-specific border fragments. These results demonstrate the commercial feasibility of improving potato cultivars by selectively adding new traits while preserving intrinsic properties, and the possibility of unambiguously identifying independent transgenic cultivars.

Base Sequence

[Measuring energy turnover in the adult swine during feeding of rations with potato starch, potatoes, beets, pressed shreds and coarse fodder as supplements to a basic ration. 1. Energy turnover and energy utilization].

25 rations with a wide variation of the content of various carbohydrates (crude fibre 36-185; water-soluble carbohydrates 25-306, starch 272-683 g/kg DM) were fed to male castrates (n = 8) of a live weight between 90 and 180 kg, and energy metabolism was measured. The rations were composed of a cereal basic ration and various feed-stuffs (potato starch-raw and soaked) potatoes (raw and steamed), sugar beet and products from it, maize pellets, dried roughage lucerne, clover, lupin, grass, rye straw meal) as supplements. The digestibility and metabolisability of the energy of the rations ranged between 88 and 62 or 86 and 60% resp. Between 2 and 4% of the consumed energy were lost in urine. The partial utilization of the metabolizable energy of the rations varied between 80% (processed supplement of potato starch) and 52% (supplement of rye straw). The rations with supplements of dried roughage were relatively well energetically utilized-between 63 and 70%. A regressively calculated difference of 17%-units was regressively calculated between the utilization of metabolizable energy of either precaecal or postileal origin.

Animal Feed

Resistance in Solanum brevidens to both potato virus Y and potato virus X may be associated with slow cell-to-cell spread.

A series of experiments was carried out to investigate the nature of the resistance of the wild potato species, Solanum brevidens, to potato virus X (PVX) and potato virus Y (PVY). In vitro inoculation of leaf protoplasts of S. brevidens and the virus-susceptible dihaploid S. tuberosum genotype PDH40 with PVX or PVY using polyethylene glycol showed that protoplasts of both species were similar in susceptibility. However, examination of protoplasts prepared from the leaves of S. tuberosum and S. brevidens inoculated 2 to 5 weeks earlier showed that the percentage of PVX- and PVY-infected leaf cells of S. tuberosum were, respectively, 45- to 100-fold and about 1000-fold greater than the percentage of infected leaf cells of S. brevidens. These results suggest that resistance in S. brevidens to both PVX and PVY could be associated with slow cell-to-cell spread rather than with slow virus replication.

Disease Susceptibility

[Methodological hints on the gas chromatographic determination of kelevan and kepone in potatoes, potato foliage and soil].

The gas chromatographic determination of Kelevan, an ethyllaevulinic acid derivative of Kepone, requires its conversion to Kepone. Thermal decomposition in the injection block of the gas chromatograph at 45 degrees C. has proved to be well suited for routine work. The sample is extracted with acetonitrile; the extract is purified and concentrated under reduced pressure. The separation of Kelevan from Kepone already metabolically produced is possible by column chromatography (silica gel; elution with benzene and 5% acetone in benzene). The determination of pre-existent Kepone and of Kelevan (after thermal conversion to Kepone) was achieved by means of a Pye-Unicam gas chromatograph. The limits of detection are 0.002 p.p.m. for Kepone, and 0.003 p.p.m. for Kelevan. The recovery of Kelevan and Kepone added to control samples (for every 10 determinations with different amounts added to potatoes, potato foliage and soil samples) ranges from 63 to 88%; the standard derivation, from +/- 1.7 to 7.3%.

Chlordecone

RFLP mapping on potato chromosomes of two genes controlling extreme resistance to potato virus X (PVX).

Two different chromosomal locations of major genes controlling extreme resistance to potato virus X (PVX) were found by restriction fragment length polymorphism (RFLP) analysis of two populations segregating for the resistance. The resistance gene Rx1 mapped to the distal end of chromosome XII, whereas Rx2 was located at an intermediate position on linkage group V in a region where reduced recombination and segregation distortion have also been observed. These linkage anomalies were due to abnormal behaviour of the chromosome contributed by the resistant parent P34. The results presented were obtained using two different strategies for mapping genes of unknown location. One approach was the use of probes revealing polymorphic loci spread throughout the genome and resulted in the mapping of Rx1. The second approach was based on the assumption of possible linkage between the resistance gene and clone-specific DNA fragments introduced from a wild potato species. Rx2 was mapped by adopting this strategy.

Blotting, Southern

Localization by restriction fragment length polymorphism mapping in potato of a major dominant gene conferring resistance to the potato cyst nematode Globodera rostochiensis.

A major dominant locus conferring resistance against several pathotypes of the root cyst nematode Globodera rostochiensis was mapped on the linkage map of potato using restriction fragment length polymorphism (RFLP) markers. The assessment of resistance versus susceptibility of the plants in the experimental population considered was based on an in vivo (pot) and an in vitro (petri dish) test. By linkage to nine RFLP markers the resistance locus Gro1 was assigned to the potato linkage group IX which is homologous to the tomato linkage group 7. Deviations from the additivity of recombination frequencies between Gro1 and its neighbouring markers in the pot test led to the detection of a few phenotypic misclassifications of small plants with poor root systems that limited the observation of cysts on susceptible roots. Pooled data from both tests provided better estimates of recombination frequencies in the linkage interval defined by the markers flanking the resistance locus.

Animals

Comparative sequence of the helper component (HC) region of potato virus Y and a HC-defective strain, potato virus C.

Potato virus C (PVC), a non-aphid transmissible strain of potato virus Y (PVY), was found to code for a protein (PVC-HC) which is similar in molecular weight and immunological reactivity to the helper component protein of PVY (PVY-HC). PVC-HC, however, was inactive with respect to its ability to effect aphid transmission of either PVC or PVY. The 5'-terminal 2.7-kb regions of PVC and PVY were sequenced. Within the HC region there was 92% nucleotide homology between the two strains; comparison of the derived amino acid sequences revealed 24 amino acid differences. Comparison of the PVC-HC sequence with that of five potyviruses revealed 2 amino acid changes which were specific to PVC-HC. These amino acids are prime targets for mutational analysis of HC activity.

Amino Acid Sequence

Replication of potato virus X RNA is altered in coinfections with potato virus Y.

Potato virus X (PVX) and potato virus Y (PVY) may coinfect tobacco to cause a classic synergistic disease. In the acute stage the disease is characterized by a dramatic increase in the accumulation of infectious PVX particles, with no corresponding increase or decrease in the accumulation of PVY. The accumulation of PVX genomic RNA and coat protein has been examined in doubly versus singly infected tobacco leaves. These experiments indicate that the levels of both viral components increase in doubly infected plants to about the same extent as the level of infectious PVX particles. The level of PVX subgenomic coat protein mRNA found associated with polyribosomes of synergistically infected plants is also increased to a similar extent. Pulse labelling experiments suggest that the increase in PVX coat protein is due to an increased rate of synthesis. The level of PVX (-) strand RNA template increases disproportionately in doubly infected tissue, to a level three times higher than that of the virion or its component parts. This result suggests that PVX/PVY synergism involves an alteration in the normal regulation of the relative levels of PVX (+) and (-) strand RNAs during viral replication.

Blotting, Northern

Comparison of antibiotic-amended potato dextrose agar and acidified potato dextrose agar as growth substrates for fungi.

Fifteen fungal species, all isolated from food, were compared for their growth abilities on potato dextrose agar acidified to pH 3.5, and on nonacidified potato dextrose agar amended with 40 ppm chlortetracycline hydrochloride. Comparisons were made at 16, 21, 26, 32, and 37 degrees C. Of the 15 species, only Penicillium expansum exhibited better growth on the acidified medium than on the nonacidified antibiotic medium, while 9 species grew better on the nonacidified antibiotic medium. Five species grew equally well on either medium.

Agar

Studies on enzymic browning of potatoes (Solanum tuberosum). III. Kinetics of potato phenoloxidase (EC 1.14.18.1 monophenol, dihydroxyphenylalanine: oxygen-oxidoreductase).

From initial velocity studies a sequential mechanism for the reactions catalysed by phenoloxidase from potatoes is indicated. The data are in accordance with an ordered addition of oxygen and phenolic substrate to the enzyme, with oxygen being the first substrate bound at thermodynamic equilibrium. The Michaelis constants for L-tyrosine, L-dopa, and chlorogenic acid are 1.4 X 10(-3), 3.3 X 10(-4), and 1.4 X 10(-4) mol/l, respectively. The dissociation constant for the enzyme-oxygen complex is about 10(-3) mol/l. In the presence of chlorogenic acid no lag phase occurs in the course of L-tyrosine oxidation. With increasing amounts of chlorogenic acid the tyrosinase activity goes through a maximum. The significance of these findings for the in vivo action of the enzyme is discussed.

Catalysis