zur Erlangung des akademischen Grades
doctor rerum agriculturarum
(Dr. rer. agr.)

vonHazem Abd El-Rahman Obiadalla Ali (M.Sc.),
geb. am 14.09.1969 in Sohag, Ägypten

Präsident der
Humboldt-Universität zu Berlin
Prof. Dr. Jürgen Mlynek

Dekan der
Landwirtschaftich-Gärtnerischen Fakultät
Prof. Dr. Uwe Jens Nagel

Gutachter:
Prof. Dr. F. Pohlheim
Prof. Dr. L. Willmitzer
Dr. J. Kossmann

Tag der mündlichen Prüfung: 10.06.2003

The work presented in this thesis was carried out between November 1999 and December 2002 at the Max-Planck-Institute für Molekulare Pflanzenphysiologie, Golm.

This Ph.D. thesis is the account of work done between November 1999 and December 2002 in the department of Prof. L. Willmitzer in the Max-Planck Institute of Molecular Plant Physiology, Golm, Germany. It is results of my own work and has not been submitted for any degree or Ph.D. at any other university.

Tabellenverzeichnis
Table 1:	Metabolite concentrations in the pericarp of 30 DAF old WT control and cp-FBP-transgenic fruits.
Table 2:	Weights and sizes of ripe tomato fruits in the WT control and cp-FBP-transgenic lines.
Table 3:	Number of flowers, fruits, fruit set and number of days to 50% flowering in the WT control and cp-FBP-transgenic lines.
Table 4:	Metabolite concentrations in the pericarp of 30 DAF old WT control and -AGP-transgenic lines.
Table 5:	Weights and sizes of ripe tomato fruits in the WT control and -AGP-transgenic lines.
Table 6:	Number of flowers, fruits, fruit set and number of days to 50% flowering in the WT control and the transgenic lines.
Table 7:	Weights and sizes of ripe tomato fruits in WT control and -GWD-transgenic lines.
Table. 8:	Number of flowers, fruit set and number of days to 50% flowering in the WT control and -GWD-transgenic lines.

Abbildungsverzeichnis
Figure 1:	The role of Fru-2,6-P2 in feedforward control of sucrose synthesis.
Figure 2:	The role of Fru-2,6-P2 in feedback control of sucrose synthesis.
Figure 3:	The predominant route of sucrose unloading and subsequent mobilization.
Figure 4:	Developmental series of tomato fruits from Micro-Tom cultivar.
Figure 5:	Starch and soluble sugar contents in pericarp and placental tissues of tomato cultivar Micro-Tom during development. (A) Starch. (B) Sucrose. (C) Fructose. (D) Glucose. Data represent the mean of five independent measurements + SE.
Figure 6:	Activities of enzymes involved in the conversion of sucrose to starch in the pericarp and placental tissues of fruit of the tomato cultivar Micro-Tom. (A) SuSy. (B) Acid invertase. (C) UDPase. (D) PGM. (E) AGPase. Data represent the mean of five independent measurements + SE.
Figure 7:	Activities of some glycolytic and clavin cycle enzymes in pericarp and placental tissues of fruit from the tomato cultivar Micro-Tom during its development.(A) HK. (B) FK. (C) FGI. (D) FBPase. (E) PPi-PFK. (F) PFK. (G) TPI. (H) G3P DH. (I) PGK. (J) Enolase. (K) PK. (L) PEP phosphatase. Data represent the mean of five independent measurements+ SE.
Figure 8:	RNA blot analysis of some plastidial transporters throughout fruit development in the tomato cultivar Micro-Tom. TPT in (A) pericarp and (B) placental tissues. Glc-6-P transporter in (C) pericarp and (D) placental tissues. ATP/ADP transporter in (E) pericarp and (F) placental tissues.
Figure 9:	Aerial parts of plants in both WT control and -cp-FBP-transgenic lines after 8 weeks growth in the glasshouse. From left to right: untransformed WT control, -cp-FBP#19, -cp-FBP#33, -cp-FBP#34 and -cp-FBP#34. The -cp-FBP plants are phenotypically identical to the untransformed WT control.
Figure 10:	FBPase activity during developmental stage (A), Western blot analysis in green (25 DAF) (B) in the pericarp of WT and -cp-FBP-transgenic lines [total soluble fruit protein (25µg) was subjected to SDS-PAGE on a 10% (w/v) gel] and FBPase activity in the leaves of WT control and -cp-FBP-transgenic lines (C). Data represent the mean of five independent measurements + SE.
Figure 11:	Starch and soluble sugar contents in pericarp of WT and -FBP-transgenic lines in tomato cultivar Moneymaker during development. (A) Starch. (B) Glucose. (C) Fructose. (D) Sucrose. Data represent the mean of five independent measurements + SE.
Figure 12:	Activities of enzymes involved in the conversion of sucrose to starch in pericarp of the WT control and cp-FBP-transgenic lines of fruit of the tomato cultivar Moneymaker. (A) SuSy. (B) UGPase. (C) PGM. (D) AGPase. Data represent the mean of five independent measurements + SE.
Figure 13:	Some 65 DAF old fruits from cp-FBP-transgenic lines (bottom) in comparison with a control fruit (above).(A) Transgenic line #19. (B) Transgenic line #33 (C) Transgenic line #34.
Figure 14:	Aerial parts of plants in both WT control and -AGP-transgenic lines after 13 weeks growth in the glasshouse. From left to right: WT control, transgenic line #2, transgenic line #7, transgenic line #11 and transgenic line #11. The -AGP plants are phenotypically identical to the untransformed WT control.
Figure 15:	AGPase activity during developmental stage (A) and Western blot analysis in green (25 DAF) (B) in the pericarp of WT control and -AGP-transgenic lines. Total soluble fruit protein (25µg) was subjected to SDS-PAGE on a 10% (w/v) gel.
Figure 16:	Starch and soluble sugar contents in the pericarp of the WT control and -AGP-transgenic lines.(A) Starch. (B) Glucose. (C) Fructose. (D) Sucrose. Data represent the mean of five independent measurements + SE in both WT control and transgenic line #7, but four independent measurements + SE in transgenic line #2 and transgenic line #11.
Figure 17:	Activities of enzymes involved in the conversion of sucrose to starch in pericarp of the WT control and -AGP transgenic lines of fruit of the tomato cultivar Moneymaker. (A) SuSy. (B) UGPase. (C) PGM. (D) FBPase. Data represent the mean of five independent measurements + SE in both WT control and transgenic line #7 and four independent measurements + SE in transgenic line #2 and transgenic line #11.
Figure 18:	Some 65 DAF old fruits from -AGP-transgenic line #7 (bottom) in comparison with the WT control fruit (above).
Figure 19:	Aerial parts of plants in both WT control and -GWD-transgenic lines after 8 weeks growth in the glasshouse. From left to right: WT control, transgenic line #16, transgenic line #17, transgenic line #20 and transgenic line #20.
Figure 20:	Immunoblot analysis of the GWD protein in (A) leaves of untransformed WT control and three selected transgenic lines [Total soluble leaf protein (15µg) was subjected to SDS-PAGE on an 8% (w/v) gel], (B) in the pericarp of the WT control and transgenic lines (25 DAF) [Total soluble fruit (pericarp) protein (30µg) was subjected to SDS-PAGE on an 8% (w/v) gel] and (C) in the pericarp of the WT control fruits between 25-70 DAF [Total soluble fruit protein (20µg) was subjected to SDS-PAGE on an 8% (w/v) gel].
Figure 21:	Starch and soluble sugar contents in the pericarp of the WT control and -GWD-transgenic lines in tomato cultivar Moneymaker during development. (A) Starch. (B) Glucose. (C) Fructose. (D) Sucrose. Data represent the mean of five independent measurements + SE in the WT control and transgenic line #17, but four independent measurements + SE in transgenic lines #16 and #20
Figure 22:	Starch and soluble sugar contents in the leaves of the WT control and transgenic tomato lines lacking the GWD protein. (A) Starch. (B) Soluble sugars. Data represent the mean of five independent measurements + SE in the WT control and transgenic line #17, but four independent measurements + SE in transgenic lines #16 and #20.
Figure 23:	Activities of enzymes involved in the conversion of sucrose to starch in the pericarp of the WT control and -GWD-transgenic lines of tomato cultivar Moneymaker.(A) SuSy. (B) UGPase. (C) PGM. (D) AGPase. (E) FBPase. Data represent the mean of five independent measurements + SE in WT control and transgenic line #17, but four independent measurements + SE in transgenic lines #16 and #20.
Figure 24:	Some 65 DAF old fruits from -GWD-transgenic lines (bottom) in comparison with the WT control fruit (above). (A) Transgenic line #16. (B) Transgenic line #17. (C) Transgenic line #20.

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