گھگھی تے پیسے
اک واری دی گل اے کہ اک ٹاہلی اتے اک گھگھی نے آہلنا پایا ہویا سی۔ اوس نے انڈے دتے جنہاں وچوں دو بہت ای سوہنے بچے نکلے۔ گھگھی بہت خوش سی۔ اوہ بچیاں لئی کول دے کھیتاں وچوں دانے اکٹھے کر کے لیاندی۔ انج کئی دن ہو گئے۔ ہن اوہدے بچے کجھ سیانے ہو گئے سن۔
اک دن اوس نوں کسے تھاں توں پیسیاں والی پوٹلی لبھی۔ اوس ایہہ پیسے اپنے آہلنے وچ لیا کے رکھ دتے۔ اک دن پنڈ دا اک منڈا گھگھی دے بچے آہلنے وچوں کڈھن لئی درخت اتے چڑھیا۔ اوس نوں پیسیاں والی پوتلی نظر آئی۔ اوس پوٹلی چکی تے تھلے اتر آیا۔ جدوں گھگھی واپس آئی تاں اوہ پوٹلی اوس نوں نظر نہ آئی تاں اوہ آکھدی اے کہ ’’کوگوں کوں کوئی بھکھا مار دا میرے پیسے لے گیا۔‘‘ اوہ منڈا تھلے کھڑا گھگھی دی ساری گل بات سن رہیا ہوندا اے۔ اوہدی گل سن کے اوس نوں بہت غصہ آندا اے۔ اوس اک وتا چک کے گھگھی دے ماریا جو اوہدے سر وچ لگا تے اوہدا سر پاٹ گیا۔ اوہ آکھدی اے کہ ’’کوگوں کوں، اسیں لال پراندے پائے نیں‘‘ اوہدا زخم ٹھیک نئیں ہوندا۔ اوہدے وچ کیڑے پے جاندے نیں تے اخیر اوہ مر جاندی اے۔ فیر اوس نوں نہوا کے کفن پایا جادنا اے تے دفن کر دتا جادنا اے۔
The article attempts to analyze the religious and legal practices inside Pakistan regarding the issue of the kitabiyyah mother and ḥaḍanah of any Muslim child. The discussion primarily addresses the major issues emphasizing kitabiyyah mother and her relatives’ right to retain the custody of the Muslim child, and duration of ḥaḍanah under the supervision of kitabiyyah mother in shari‘ah and Pakistani Family Laws. Furthermore, it argues that kitabiyyah mother is permitted to raise the Muslim child according to her faith under both Islamic and Pakistani Family Laws. The article is delimited to the opinions of four Sunni schools of thoughts, Statute law, and Case laws. Nonetheless, in-depth comparative analysis has been carried out in most persuasive way to examine the rules related to kitabiyyah mother in custody of Muslim child after divorce under both Islamic Law, and Case Laws in Pakistan. Additionally, the existing similarities and differences consequent to religious differences have also been appropriately figured out to point out plausible way forward to address prevailing schism. Contradictions between the legal practices and Islamic law need keen attention Islamic and legal scholars to carefully craft to harmonize both in the best interest of child.
The three dehydrin genes of Eucalyptus species, the dehydrin-1 (EgDHN-1), dehydrin-2 (EgDHN-2) and dehydrin-10 (EgDHN-10) were isolated from local Eucalyptus globulus species for transformation in different varieties of potato for developing abiotic stress tolerance. These three lysine rich dehydrin genes of Eucalyptus have not been reported earlier to be manipulated for genetic engineering or for transformation into a crop plant. The genes were amplified from DNA as well as mRNA (through cDNA). The amplified genes were cloned in TA cloning vector. Cloned dehydrin genes were sequenced and detailed in silico analyses were conducted. Complete sequence of EgDHN-10 gene was obtained from DNA including introns. Coding sequence from its mRNA was also complete and found 92% similar with EgDHN-10 of Eucalyptus globulus after DNA BLAST analyses. The sequencing data of other genes obtained incomplete that showed 99% and 98% homology, after DNA BLAST analyses, with Predicted Eucalyptus grandis Rab18 gene and Eucalyptus globulus EgDHN-1 gene respectively. 297bp long sequence of EgDHN-10 that codes for mature peptide was further sub-cloned in E. coli expression vector pET30a. The sequence was submitted to the NCBI (MG948256.1) and it had extra integration of five electrically charged amino acids (two positively charged Histidine, one negatively charged Glutamate, and two negatively charged Aspartate residues) and one Lysine (K). The presence of one extra K residue is very important for enhanced stress tolerance activity of the transformed dehydrin protein due to its role in functions of K-segments. Presence of more charged amino acids in the binding surface of dehydrin protein enhances its capability to bind with other macromolecules, reactive oxygen species and free metal ions.IPTG induced successful expression of EgDHN-10, cloned in pET30a, was detected in E. coli BL21 ii DE3 strain by protein dot blot analysis. After getting excellent expression of Eucalyptus globulus EgDHN-10 gene outside Eucalyptus cell’s environment, the gene was sub-cloned in pCAMBIA 1301 plant binary vector and transformed into Agrobacterium tumefascience. The nodal regions of three potato cultivars, the Cardinal, Sante and Lady Rosetta, were infected with transformed Agrobacterium (through co-cultivation) carrying EgDHN-10. Putative transgenic plants were first screened by GUS assay and PCR. The successful integration of EgDHN-10 gene in potato genome was confirmed by DNA dot blot analysis. The EgDHN-10 gene’s expression was confirmed from potato mRNA by PCR and real time quantitative PCR. In some plants up to 12 folds DHN 10 expression was observed through qPCR. The acquired abiotic stress tolerance was quantified in stressed transformed potato plants by ion leakage assay (ILA) of cell rupture to select the stable transgenic plants. The stress tolerant transformed plants of Cardinal, Sante and Lady Rosetta were maintained in plant tissue culture setup and pots. The transgenic Sante variety was multiplied and maintained in CAMB trial field in an experiment of growth promoting effect of biofertilizer on trial field plants. The stress tolerant Sante plants showed significant difference over control plants in studied plant growth parameters like plant height, maximum root length, number of roots and number of multiple shoots formation, number and weight of tuber formation.