Genetic and Molecular Analysis of Transgenic Rice cv. Rojolele Expressing Lactoferrin

Abstract

In a previous study, human lactoferrin gene have introduced into Javanica rice cv. Rojolele by Agrobacterium-mediated transformation. Lactoferrin (LF) is an 80 kDa iron-binding glycoprotein that has been proposed to have many biological roles such as protection against microbial and virus infection. This study aims to analyze the integration and level of lactoferrin gene expression of transgenic rice cv. Rojolele. The study also aims to examine the genetic character of transgenic rice expressing recombinant lactoferrin. Stability expression of recombinant lactoferrin transgenic rice seeds over generations were analyzed by ELISA, while the integration stability of recombinant hLF gene in transgenic plants performed by PCR. The mitotic time, cell cycle and chromosome characterization of transgenic and non-transgenic rice cv. Rojolele were determined. Chromosome characterization of the trangenic and non transgenic rice cv. Rojolele was investigated to determine the genetic variation. All of the above efforts were aimed to evaluate the genetically engineered rice containing recombinant lactoferrin as a nutraceutical food. The results showed that the expression was stable through three consecutive generations. The expression of the hLF gene increased during grain-filling period. The active time of mitotic cells of transgenic rice rojolele was longer than the cells of non-transgenic rice. In addition, the cycle cell of transgenic and non-transgenic rojolele contained prophase, prometaphase, metaphase, anaphase, telophase and interphase. The result showed that all of the transgenic lines had diploid (2n) chromosome number = 24.

Rojolele: a Premium Aromatic Rice Variety in Indonesia

“…Lactoferrin gene have been introduced to Rojolele by Agrobacterium-mediated transformation [50]. Lactoferrin plays an important role as a nutraceutical food due its function against virus and microbial infection.…”

Agronomic characteristics and genetic relationship of putative transgenic rice lines of cv. Fatmawati with the Glu-1Dx5 transgene

“…Twenty-seven (46%), out of 59 independent transgenic events are tetraploid (2n=2x= 28) or aneuploid around the tetraploid level (i.e., 26, 27, 29 and 30 chromosomes;Choi et al 2000). However, all transgenic rice lines that express human lactoferrin have diploid (2n=2x= 24 chromosomes; Rachmawati et al 2014). Contradiction results between these two transgenic experiments can be attributed to the differences in plant type, callus age, callus culture condition, transformation method, insertional effect, retrotransposon activation, and other possible cause for this condition.…”

Rojolele: A Premium Aromatic Rice Variety in Indonesia

“…Lactoferrin gene have been introduced to Rojolele by Agrobacterium-mediated transformation (Rachmawati et al, 2014). Lactoferrin plays an important role as a nutraceutical food due its function against virus and microbial infection.…”